CA2886397A1 - Methods and compositions relating to next generation sequencing for genetic testing in alk related cancers - Google Patents

Abstract

Disclosed are methods, assays, and compositions for detecting the presence of a kinase inhibitor resistance. The disclosed method and primer panels work with any method for detecting nucleic acid variation in a sample including, but not limited to next generation sequencing.

Description

METHODS AND COMPOSITIONS RELATING TO NEXT GENERATION
SEQUENCING FOR GENETIC TESTING IN ALKRELATED CANCERS
BACKGROUND
Mutations of anaplastic lymphoma kinase (A L K) gene are thought to be involved in the development of subsets of numerous cancers including i) non-small cell lung carcinoma (NSCLC); ii) diffuse large B-cell lymphoma; iii) esophageal squamous cell carcinoma; iv) anaplastic large-cell lymphoma (ALCL); v) neuroblastoma (a childhood cancer that arises from the developing peripheral nervous system); and vi) the sarcomas known as inflammatory myofibroblastic tumors (IMTs). Patient outcomes with many of these malignancies are poor, due in part to the late detection of the cancers because of the lack of efficient clinical diagnostic methods. Early detection and diagnosis of ALK-mediated cancers dramatically increases survival rates within the patient population; as an example, early detection of ALK-positive anaplastic large-cell lymphoma can result in survival rates of up to 83% whereas late detection is associated in some instances with survival of only 50% of the patient population.
The critical role of deregulated ALK signaling as a driver of subsets of NSCLC, ALCL, and other ALK-dependent cancer types has been validated in clinical trials, with dramatic anti-tumor efficacy observed in response to the ALK small-molecule inhibitor crizotinib (XALKORT , Pfizer; approved by the US FDA in August 2011).
Unfortunately, despite the marked anti-tumor responses to XALKORT seen in patients with ALK-driven tumors, most patients eventually experience progression of their cancer as a consequence of treatment resistance. For example, the median duration of progression-free survival in patients with ALK-positive NSCLC treated with Xalkori is only about 10 months. What is needed are assays the can efficiently and reliably detect kinase inhibitor-resistance mutations and therefore predict which members of a patient population is likely to develop kinase inhibitor resistance. Additionally as new generations of small-molecule inhibitors are developed, also need is a clinically applicable diagnostic test to identify resistance mutations in the ALK kinase domain and therefore to guide the rational use of these small-molecule inhibitors for the treatment of ALK-driven cancers that have lost their responsiveness to 1st-generation inhibitor therapy. Moreover, once several ALK small-molecule inhibitors are approved for clinical use, optimal management of patients with ALK-driven tumors will require screening for de novo inhibitor resistance mutations by healthcare providers treating newly diagnosed patients in order to assess their inhibitor sensitivity and choose the best ALK inhibitor drug(s) for personalized therapy.
BRIEF SUMMARY
The methods, assays, and compositions disclosed herein relate to the field of detection or diagnosis of mutations that confer resistance to kinase inhibitors of a disease or condition such as cancer. In one aspect, the kinase inhibitors or ALK kinase inhibitors.
Also disclosed herein are methods and assays for assessing the susceptibility or risk for developing resistance to an inhibitor, wherein the disease or condition is a cancer associated with expression of the ALK gene. It is understood and herein contemplated that the methods disclosed herein allow for rapid and sensitive detection of nucleic acid expression of mutations in ALK.
In another aspect, disclosed herein are kinase inhibitor resistance panels comprising one or more primer sets from each of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
In accordance with the purpose(s) of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to an ALK kinase inhibitor resistance panel. In particular, the invention, in one aspect, relates to an ALK kinase inhibitor resistance panel comprising one or more primer sets for detecting the presence of a mutation in a gene that will confer resistance to the ALK kinase inhibitor.
Additional advantages of the disclosed methods and compositions will be set forth in part in the description which follows, and in part will be understood from the description, or may be learned by practice of the disclosed method and compositions. The advantages of the disclosed methods and compositions will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be

2

3 understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as claimed.
BRIEF DESRIPTION OF THE DRAWINGS
Figure 1 shows XALKORIO-resistance mutations identified in patient specimens.
The figure depicts the XALKORIO-resistance mutations in the ALK kinase domain identified to date in patient cancer specimens.
DETAILED DESCRIPTION
Before the present compounds, compositions, articles, devices, and/or methods are disclosed and described, it is to be understood that they are not limited to specific synthetic methods or specific recombinant biotechnology methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary.
It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.
As used in the specification and the appended claims, the singular forms "a,"
"an"
and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a pharmaceutical carrier" includes mixtures of two or more such carriers, and the like.
Ranges can be expressed herein as from "about" one particular value, and/or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value.
Similarly, when values are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as "about"
that particular value in addition to the value itself For example, if the value "10" is disclosed, then "about 10" is also disclosed. It is also understood that when a value is disclosed that "less than or equal to" the value, "greater than or equal to the value" and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value "10" is disclosed the "less than or equal to 10"as well as "greater than or equal to 10" is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point "10" and a particular data point 15 are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15.
In this specification and in the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings:
"Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.
An "increase" can refer to any change that results in a larger amount of a composition or compound, such as an amplification product relative to a control. Thus, for example, an increase in the amount in amplification products can include but is not limited to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 9u,-so ,/0 , 100%, 150%, 200%, 250%, 300%, 350%, 400%, 450%, 500%, 550%, 600%, 700%, 800%, 900%, 1000%, 1500%, 2000%, 2500%, 3000%, 3500%, 4000%, 4500%, or 5000% increase. It is further contemplated herein that the detection an increase in expression or abundance of a DNA, mRNA, or protein relative to a control necessarily includes detection of the presence of the DNA, mRNA, or protein in situations where the DNA, mRNA, or protein is not present in the control.
"Obtaining a tissue sample" or "obtain a tissue sample" means to collect a sample of tissue either from a party having previously harvested the tissue or harvesting directly from a subject. It is understood and herein contemplated that tissue samples obtained directly

4 from the subject can be obtained by any means known in the art including invasive and non-invasive techniques. It is also understood that methods of measurement can be direct or indirect. Examples of methods of obtaining or measuring a tissue sample can include but are not limited to tissue biopsy, tissue lavage, blood collection, aspiration, tissue swab, spinal tap, magnetic resonance imaging (MRI), Computed Tomography (CT) scan, Positron Emission Tomography (PET) scan, and X-ray (with and without contrast media).
It is further understood that a "tissue" can include, but is not limited to any grouping of one or more cells or analytes to be used in a an ex vivo or in vitro assays. Such tissues include but are not limited to blood, saliva, sputum, lymph, cellular mass, and tissue collected from a biopsy.
Kinase Inhibitor Resistant Panels In one aspect, disclosed herein are kinase inhibitor resistance panels such as, for example, an ALK kinase inhibitor panel. Kinase inhibitors are known in the art and have found use in the treatment of, amongst other things, the treatment of cancer.
For example, cancers involving the overexpression or fusion of Analplastic Lymphoma Kinase can be treated through the use of a kinase inhibitor. Kinase inhibitors are known in the art and include, but are not limited to crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib. Thus, in one aspect, disclosed herein are kinase inhibitor resistance panels for detecting susceptibility or resistance to treatment in a subject to a kinase inhibitor comprising crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, or Vemurafenib.
Unfortunately, mutations in the ALK sequence and other genes, such as, BRAF, KIT, KRAS, and EGFR can lead to kinase inhibitor resistance. These mutations can comprise any

5 of the mutations to ALK, KIT, BRAF, KRAS, or EGFR listed in Tables 2, 3, 4, 5, or 6.
Accordingly, in a further aspect, disclosed herein are kinase inhibitor panels comprising one or more primer sets that selectively hybridize and can be used to amplify one of the genes selected from group of genes comprising KRAS(SEQ ID NO: 7718), BRAF(SEQ ID NO:
7717), EGFR (SEQ ID NO: 7716), ALK (SEQ ID NO: 7714 and SEQ ID NO: 7717 (cDNA)), and KIT. In one aspect, the kinase inhibitor resistance panel disclosed herein can comprise one or more primer set(s) that hybridizes and amplifies nucleic acid from exon 1 (SEQ ID NOs: 4601-4880 and 7181-7230) exon 2 (SEQ ID NOs: 4881-5200 and 7231-7326) or both exons 1 and 2 (SEQ ID NOs: 7327-7610) of KRAS; exon 18 (SEQ ID
NOs:
1641-1760 and 5819-5934), exon 19 (SEQ ID NOs: 1761-1880), exon 20 (SEQ ID
NOs:
1881-2000 and 5934-6042), exon 21 (SEQ ID NOs: 2001-2120 and 6043-6150), exon (SEQ ID NOs: 2121-2240, 2321-2360, and 2401-2440), exons 18 and 19 (SEQ ID
NOs:
2241-2280), exons 18, 19, and 20 (SEQ ID NOs: 6151-6274), exons 20 and 21 (SEQ
ID
NOs: 2281-2320 and 6275-6388), or exons 18, 19, 20, and 21 (SEQ ID NOs: 2361-and 6389-6524) of EGFR; exon 8 (SEQ ID NOs: 2441-2800), exon 9 (SEQ ID NOs:

3120), exon 10 (SEQ ID NOs: 3201-3360), exon 11 (SEQ ID NOs: 3361-3480), exon (SEQ ID NOs: 3481-3640), exon 13 (SEQ ID NOs: 3641-3800), exon 17 (SEQ ID NOs:

4241-4600), exon 8 and 9 (SEQ ID NOs: 2801-2840), exons 9 and 10 (SEQ ID NOs:

3160), exons 9, 10, and 11 (SEQ ID NOs: 3161-3200); exons 10 and 11 (SEQ ID
NOs:
3801-3960), exons 12 and 13 (SEQ ID NOs: 3961-4120), or exons 10, 11, 12, and 13 (SEQ
ID NOs: 4121-4240) of KIT; exons 10 and 11 (SEQ ID NOs: 6525-6832) or exons 13, 14, or 15 (SEQ ID NOs: 66833-7180) of BRAF , and/or exon 21 (SEQ ID NOs: 1-160), exon 22 (SEQ ID NOs: 401-560), exon 23 (SEQ ID NOs: 561-840 and 5311-5446), exon 24 (SEQ
ID NOs: 921-1240), exon 25 (SEQ ID NOs: 1241-1600), exons 21 and 22 (SEQ ID
NOs:
161-400 and 5201-5310), exons 21, 22, and 23 (SEQ ID NOs: 841-920), exons 24 and 25 (SEQ ID NOs: 1601-1640 and 5447-5576), or exons 21, 22, 23, 24, and 25 (SEQ ID
NOs:
5577-5818) of ALK. As disclosed herein "primer set" refers to a forward and reverse primer pair (i.e., a left and right primer pair) that can be used together to amplify a given region of

6 a nucleic acid (e.g., DNA, RNA, or cDNA) of interest. Thus, panels with multiple primer sets include multiple primer pairs. It is understood and herein contemplated that some primer sets may have a common forward or reverse primer and thus have an odd number of primers.
It is further understood and herein contemplated that the disclosed kinase inhibitor resistant panels can comprise a single primer sets that hybridizes to a single gene, region, or exon of a gene selected from the group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, a single primer sets for KRAS, BRAF, EGFR, ALK, or KIT); multiple primer sets that hybridize to a single gene, region, or exon of a gene selected from the group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, one or more primer sets for KRAS, BRAF, EGFR, ALK, or KIT); multiple primer sets comprising a single primer set that specifically hybridize to a single gene, region, or exon for each of the genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, a single primer set for each of KRAS, BRAF, EGFR, ALK, and/or KIT); or multiple primer sets comprising where in there is more than one primer set for each gene, region or exon for each of the genes selected from the group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT (i.e, one or more primer sets for each of KRAS, BRAF, EGFR, ALK, and/or KIT). Thus, it is contemplated herein that the kinase inhibitor panel can comprise primer sets that recognize and specifically hybridize to a gene, region, or exon, of one or combination of the gene selected from the group consisting of KRAS, BRAF, EGFR, ALK, and KIT. For example, the panel can comprise primer sets that hybridize to a gene, region, or exon of KRAS, BRAF, EGFR, ALK, or KIT;
KRAS and BRAF; KRAS and EGFR; KRAS and ALK; KRAS and KIT; BRAF and EGFR;
BRAF and KIT; BRAF and ALK; EGFR and ALK; EGFR and KIT; ALK and KIT; KRAS, BRAF, and EGFR; KRAS, BRAF, and ALK; KRAS, BRAF, and KIT; KRAS, EGFR, and ALK; KRAS, EGFR, and KIT; KRAS, ALK, and KIT; BRAF, EGFR, and ALK, BRAF, EGFR, and KIT; BRAF, ALK, and KIT; EGFR, ALK, and KIT; KRAS, BRAF, EGFR, and ALK; KRAS, BRAF, EGFR, and KIT, BRAF, EGFR, ALK, and KIT; and KRAS, BRAF, EGFR, ALK, and KIT.

7 For example, the primer or primer sets in the kinase inhibitor resistance panel can detect any of the mutations in Tables 2-6. In another aspect, the primers or primer sets used in the inhibitor resistance panel can comprise one or more of the primers or primer sets listed in Tables 7-14 as disclosed herein and/or probes listed in Table 15 (i.e., SEQ ID NOs:
7611-7613).
Methods of detecting the presence of a kinase inhibitor resistant cancer The disclosed kinase inhibitor resistant panels, in one aspect, contain primers or primer sets for the detection of mutations that confer kinase inhibitor resistance. Thus, in another aspect disclosed herein are methods and assays for the detection of kinase inhibitor resistant forms of an ALK-related cancer. For example, disclosed herein are methods and assays for the detection of kinase inhibitor resistance, such as, for example ALK kinase inhibitor resistance, comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor. In one aspect, the mutation can be a nucleic acid mutation in ALK, EGFR, KRAS, BRAF, or KIT. For example, the mutation can be any mutation listed in Tables 2-6.
In a further aspect, the disclosed methods and assays for detection of kinase inhibitor resistance can comprise performing next generation sequencing using a kinase inhibitor resistant panel as disclosed herein which comprises a primer or primer set that hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS; exon 18, 19, 20, 21 or 22 of EGFR;
exon 8, 9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11,13, 14, or 15 of BRAF , and/or exon 21, 22, 23, 24, or 25 of ALK. For example, the primer or primer set can comprise any of the primers or primer sets disclosed in Tables 7-14. Thus, disclosed herein are methods wherein the one or more primer set(s) that hybridizes and amplifies nucleic acid from exon 1 (SEQ ID NOs: 4601-4880 and 7181-7230) exon 2 (SEQ ID NOs: 4881-5200 and 7231-

8 7326) or both exons 1 and 2 (SEQ ID NOs: 7327-7610) of KRAS; exon 18 (SEQ ID
NOs:
1641-1760 and 5819-5934), exon 19 (SEQ ID NOs: 1761-1880), exon 20 (SEQ ID
NOs:
1881-2000 and 5934-6042), exon 21 (SEQ ID NOs: 2001-2120 and 6043-6150), exon (SEQ ID NOs: 2121-2240, 2321-2360, and 2401-2440), exons 18 and 19 (SEQ ID
NOs:
2241-2280), exons 18, 19, and 20 (SEQ ID NOs: 6151-6274), exons 20 and 21 (SEQ
ID
NOs: 2281-2320 and 6275-6388), or exons 18, 19, 20, and 21 (SEQ ID NOs: 2361-and 6389-6524) of EGFR; exon 8 (SEQ ID NOs: 2441-2800), exon 9 (SEQ ID NOs:

3120), exon 10 (SEQ ID NOs: 3201-3360), exon 11 (SEQ ID NOs: 3361-3480), exon (SEQ ID NOs: 3481-3640), exon 13 (SEQ ID NOs: 3641-3800), exon 17 (SEQ ID NOs:
4241-4600), exon 8 and 9 (SEQ ID NOs: 2801-2840), exons 9 and 10 (SEQ ID NOs:

3160), exons 9, 10, and 11 (SEQ ID NOs: 3161-3200); exons 10 and 11 (SEQ ID
NOs:
3801-3960), exons 12 and 13 (SEQ ID NOs: 3961-4120), or exons 10, 11, 12, and 13 (SEQ
ID NOs: 4121-4240) of KIT; exons 10 and 11 (SEQ ID NOs: 6525-6832) or exons 13, 14, or 15 (SEQ ID NOs: 66833-7180) of BRAF, and/or exon 21 (SEQ ID NOs: 1-160), exon 22 (SEQ ID NOs: 401-560), exon 23 (SEQ ID NOs: 561-840 and 5311-5446), exon 24 (SEQ
ID NOs: 921-1240), exon 25 (SEQ ID NOs: 1241-1600), exons 21 and 22 (SEQ ID
NOs:
161-400 and 5201-5310), exons 21, 22, and 23 (SEQ ID NOs: 841-920), exons 24 and 25 (SEQ ID NOs: 1601-1640 and 5447-5576), or exons 21, 22, 23, 24, and 25 (SEQ ID
NOs:
5577-5818) of ALK.
It is understood that the disclosed methods can further comprise synthesizing cDNA
from the nucleic acid extracted from a tissue sample before detection of a mutation in ALK, EGFR, KRAS, BRAF, or KIT. Thus, in one aspect, disclosed herein are methods for detecting kinase inhibitor resistance in a cancer in a subject, for example ALK kinase inhibitor resistance, comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); synthesixing cDNA from the tissue sample, and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid

9 sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.
It is further understood and herein contemplated that the subject of the disclosed methods can be a subject that has been previously diagnosed with a cancer including but not limited to inflammatory breast cancer, non-small cell lung carcinoma, esophageal squamous cell carcinoma, colorectal carcinoma, Inflammatory myofibroblastic tumor, familial and sporadic neuroblastoma. In yet another aspect, the subject has been previously diagnosed with a cancer that results from ALK, ROS1, RET, DEPDC1 overexpression, dysregulation, or fusion. Examples of such fusions include but are not limited to nucleophosmin-ALK
(NPM-ALK), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP
cyclohydrolase (ATIC-ALK), clathrin heavy chain-ALK (CLTC-ALK), kinesin-1 heavy chain gene-ALK (KIF5B-ALK); Ran-binding protein 2-ALK (RANBP2-ALK), SEC31L1-ALK, tropomyosin-3-ALK (TPM3-ALK), tropomyosin-4-ALK (TPM4-ALK), TRK-fused gene(Large) -ALK (TFGL-ALK), TRK-fused gene(Small) -ALK (TFGs-ALK), CARS-ALK, EML4-ALK, 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP
cyclohydrolase-ALK (ATIC-ALK), AL017-ALK, moesin-ALK (MSN-ALK), non-muscle myosin heavy chain gene-ALK (MYH9-ALK), and TRK-fused gene(Extra Large) -ALK
(TFGxL¨ALK). In a further aspect, the present methods could not only be used to diagnose a kinase inhibitor resistant cancer , but diagnose the cancer itself as the subject with a kinase inhibitor resistant cancer would necessarily not only have a cancer, but have a kinase related cancer such as those disclosed herein.
Therefore, in one aspect, disclosed herein are methods for the detection of kinase inhibitor resistance comprising obtaining a tissue sample from a subject with a cancer and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample using one or more primer sets or primer panels with primer sets that specifically hybridizes to one or more of the genes selected from the group consisting of ALK, KRAS, EGFR, KIT, and BRAF, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.
Also disclosed are methods, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 18, 19, 20, 21 or 22 of EGFR, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 21, 22, 23, 24, or 25 of ALK, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 8, 9, 10, 11, 12, 13, or 17 of KIT, and/or wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 10, 11, 13, 14, or 15 of BRAF.
In one aspect, disclosed are methods, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of Tables 10 and/or 14 (SEQ ID
NOs: 4601-5200 and 7181-7610); wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of Tables 8 and/or 12 (1641-2440 and 5819-6524); wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of Tables 7 and/or 11 (SEQ ID NOs: 1-1640 and 5201-5818);
wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of Table 9 ( SEQ ID NOs: 2441-4600); and/or wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of Table 13 (SEQ ID NOs:
6525-7180).
In one aspect are methods comprising the use of a kinase inhibitor resistance panel, wherein the panel comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
In another aspect, disclosed are methods wherein the panel comprises one or more primer sets for 2, 3, 4, of all 5 of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.
Also disclosed are methods, wherein the kinase inhibitor is selected from the group consisting of crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib.
Methods, assays, and primer panels for assessing the suitability of ALK
directed treatments Though not wishing to be bound by current theories, it is believed that inhibition of these over-expression or aberrant expressions of ALK with small-molecule drug candidates abrogates related abnormal cell proliferation and promotes apoptosis in ALK-related tumor cell lines. Furthermore, both preclinical animal models and the early clinical experience with these inhibitors indicate that ALK small-molecule inhibitors not only possess marked antitumor activity against ALK-related cancers but are also very well tolerated with no limiting target-associated toxicities. Therefore, such small molecules can be used to treat ALK-driven cancers.
However, the presence of a mutation in one of the genes associated with an ALK-related cancer can confer resistance to treatment with a kinase inhibitor, such as an ALK
kinase inhibitor. Nevertheless, knowledge of the presence of said mutation can still be useful to the practicing physician in assessing the suitability of a treatment or prescribing a particular treatment regimen. For example, the presence of a mutation in a gene which confers kinase inhibitor resistance, such as, for example, ALK kinase inhibitor resistance, can inform the skilled artisan to choose a particular kinase inhibitor over another due to the presence of a mutation affecting one kinase inhibitor and not the other.
Alternatively, the presence of a mutation can inform the physician to discontinue the course of treatment with one kinase inhibitor due to detection of kinase inhibitor resistance and select a different kinase inhibitor to which the patient is not yet resistant. Accordingly, disclosed herein are methods and assays for assessing the suitability of an ALK inhibitor treatment for a cancer, for example, NSCLC, in a subject comprising performing high throughput sequencing on nucleic acid from a tissue sample from the subject; wherein the presence of a mutation in ALK, EGFR, BRAF, KRAS, or KIT indicates a cancer that comprises resistance to an ALK

kinase inhibitor. In one aspect, disclosed herein are methods and assays for assessing a subject's suitability for treatment with a kinase inhibitor comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); detecting the presence of a mutation through sequencing or other nucleic acid detection technique for the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor and therefore continued use of an inhibitor to which the cancer has become resistant or to which the cancer is already resistant should be discontinued in favor of a cancer to which resistance has not developed.
It is understood and herein contemplated that any of the disclosed nucleic acid sequencing techniques disclosed herein can be used in these methods. Thus, disclosed herein are methods and assays assessing the suitability of an ALK kinase inhibitor treatment for an ALK related cancer in a subject comprising conducting high throughput sequencing (also known as next generation sequencing) on nucleic acid such as mRNA or DNA
from a tissue sample from the subject; wherein the sequencing reaction reveals the nucleic acid sequence for one or more exons of KIT, BRAF, KRAS, EGFR, and ALK; and wherein the presence of one or more mutations in KIT, BRAF, KRAS, EGFR, and/or ALK
indicates the presence of kinase inhibitor resistance. The mutations can occur in any exon of KIT, BRAF, KRAS, EGFR, and ALK. Thus, for example, the mutations can occur in and therefore the primers or primer sets can hybridize to exon 1 or 2 of KRAS; exon 18, 19, 20, 21 r 22 of EGFR; exon 8,9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11, 13, 14, or 15 of BRAF, and/or exon 21, 22, 23, 24, or 25 of ALK. In one aspect, the mutation can comprise any one or more of the mutations listed in Tables 2-6. It is further understood that the disclosed methods and assays can further comprise any of the primers disclosed herein in Tables 7-14 or probes listed in Table 15 and utilize the multiplexing PCR techniques disclosed.
In another aspect, two or more of the disclosed primers and primer sets can comprise a primer panel can be used in methods and assays for the assessment of the suitability of a kinase inhibitor for the treatment of a subjects' cancer. In one aspect, the primer panel comprises one or more primers that can detect a nucleic acid mutation in ALK, BRAF, EGFR, KRAS, or KIT. In a further aspect, the primers or primer sets that hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS; exon 18, 19, 20, 21 or 22 of EGFR; exon 8,9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11, 13, 14, or 15 of BRAF , and/or exon 21, 22, 23, 24, or 25 of ALK. In another aspect, the disclosed primer panel can comprise any primer or primer set which detects one or more of the mutations found in Tables 2-6.
For example, the primer or primer set can comprise any of the primers or primer sets disclosed in Tables 7-14.
In another aspect, knowledge of kinase inhibitor resistant cancer can be used to screen for a drug that is not a kinase inhibitor. Thus, in one aspect, disclosed herein are methods of screening for a drug to treat a subject with a cancer comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject has a cancer is resistant or will become resistant to a kinase inhibitor, and contacting a tissue sample from subject with a cancer with an agent; wherein an agent that inhibits or reduces the growth or development of a kinase inhibitor resistant cancer is not a kinase inhibitor. The disclosed methods can further comprise the sue of the kinase inhibitor resistant panels disclosed herein or any of the primers, primer sets or probes disclosed herein. The methods can also further comprise the treatment of a subject with a kinase inhibitor resistant cancer with an agent that is identified in the method as not being a kinase inhibitor or discontinuing treatment in a subject with kinase inhibitor resistant cancer with an agent that has been found to be a kinase inhibitor.
Methods of identifying subjects for participation in clinical trials to screen for new cancer treatments.

In one aspect, it is contemplated herein that the identification of individuals with a kinase inhibitor resistant cancer can be useful for establishing clinical trials to screen for drugs that can be used to treat individuals with kinase inhibitor resistant cancers. Thus, in one aspect, disclosed herein are methods for identifying a subject for screening for a drug that can treat a cancer in a subject with a kinase inhibitor resistant cancer, for example ALK
kinase inhibitor resistance, comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject has a cancer is resistant or will become resistant to a kinase inhibitor and the subject can be used in trials to screen for a drug to which a kinase inhibitor resistant subject will respond.
In one aspect, the mutation can be a nucleic acid mutation in ALK, EGFR, KRAS, BRAF, or KIT.
For example, the mutation can be any mutation listed in Tables 2-6. In one aspect, said methods can further comprise synthesizing cDNA from the tissue sample of the subject.
It is understood and herein contemplated that the disclosed methods can be used in conjunction with any of the kinase inhibitor resistant panels, primer sets, or probes disclosed herein. For example, the disclosed methods can be performed using a primer or primer set that hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS; exon 18, 19, 20, 21 or 22 of EGFR; exon 8, 9, 10, 11, 12, 13, or 17 of KIT; exon 10, 11, 13, 14, or 15 of BRAF , and/or exon 21, 22, 23, 24, or 25 of ALK. For example, the primer or primer set can comprise any of the primers or primer sets disclosed in Tables 7-14. Thus, disclosed herein are methods wherein the one or more primer set(s) that hybridizes and amplifies nucleic acid from exon 1 (SEQ ID NOs: 4601-4880 and 7181-7230) exon 2 (SEQ ID NOs:

5200 and 7231-7326) or both exons 1 and 2 (SEQ ID NOs: 7327-7610) of KRAS;
exon 18 (SEQ ID NOs: 1641-1760 and 5819-5934), exon 19 (SEQ ID NOs: 1761-1880), exon (SEQ ID NOs: 1881-2000 and 5934-6042), exon 21 (SEQ ID NOs: 2001-2120 and 6043-6150), exon 22 (SEQ ID NOs: 2121-2240, 2321-2360, and 2401-2440), exons 18 and (SEQ ID NOs: 2241-2280), exons 18, 19, and 20 (SEQ ID NOs: 6151-6274), exons 20 and 21 (SEQ ID NOs: 2281-2320 and 6275-6388), or exons 18, 19, 20, and 21 (SEQ ID
NOs:
2361-2400 and 6389-6524) of EGFR; exon 8 (SEQ ID NOs: 2441-2800), exon 9 (SEQ
ID
NOs: 2841-3120), exon 10 (SEQ ID NOs: 3201-3360), exon 11 (SEQ ID NOs: 3361-3480), exon 12 (SEQ ID NOs: 3481-3640), exon 13 (SEQ ID NOs: 3641-3800), exon 17 (SEQ
ID
NOs: 4241-4600), exon 8 and 9 (SEQ ID NOs: 2801-2840), exons 9 and 10 (SEQ ID
NOs:
3121-3160), exons 9, 10, and 11 (SEQ ID NOs: 3161-3200); exons 10 and 11 (SEQ
ID
NOs: 3801-3960), exons 12 and 13 (SEQ ID NOs: 3961-4120), or exons 10, 11, 12, and 13 (SEQ ID NOs: 4121-4240) of KIT; exons 10 and 11 (SEQ ID NOs: 6525-6832) or exons 13, 14, or 15 (SEQ ID NOs: 66833-7180) of BRAF, and/or exon 21 (SEQ ID NOs: 1-160), exon 22 (SEQ ID NOs: 401-560), exon 23 (SEQ ID NOs: 561-840 and 5311-5446), exon 24 (SEQ ID NOs: 921-1240), exon 25 (SEQ ID NOs: 1241-1600), exons 21 and 22 (SEQ
ID
NOs: 161-400 and 5201-5310), exons 21, 22, and 23 (SEQ ID NOs: 841-920), exons 24 and 25 (SEQ ID NOs: 1601-1640 and 5447-5576), or exons 21, 22, 23, 24, and 25 (SEQ
ID
NOs: 5577-5818) of ALK.
Methods of detecting a kinase inhibitor resistance in an ALK-related cancer In another aspect, the disclosed methods and assays relate to the detection or diagnosis of the presence of a kinase inhibitor resistance, such as, for example, ALK kinase inhibitor resistance, in a disease or condition such as a cancer and methods and assays for the determination of susceptibility or resistance to therapeutic treatment for a disease or condition such as a cancer in a subject comprising detecting the presence or measuring the expression level of nucleic acid (for example, DNA, mRNA, cDNA, RNA, etc) through the use of next generation sequencing (NGS) from a tissue sample from the subject;
wherein the presence of a mutations in the nucleic acid code of the KIT, BRAF, KRAS, EGFR, or ALK
gene or the ALK gene portion of an ALK fusion construct indicates the presence of a cancer that is resistant to a kinase inhibitor. In one aspect, the cancer is associated with amplification, overexpression, nucleic acid variation, truncation, or gene fusion of ALK. It is understood, that the kinase inhibitor resistance panels disclosed herein can be used to perform said methods and the detection of one or more of the mutations in Tables 2-6 indicates the presence of kinase inhibitor resistance. In one aspect, the disclosed methods can further comprise discontinuing use of a kinase inhibitor to treat a cancer in a subject that has been identified with a kinase inhibitor resistant cancer. In another embodiment, the disclosed methods can further comprise treating a subject with a kinase inhibitor resistant cancer with a chemotherapeutic that is not a kinase inhibitor. Thus, in one aspect, disclosed herein are methods of treating a subject with a kinase inhibitor resistant cancer (such as, for example, an ALK kinase inhibitor resistant cancer) comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers);
conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the presence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject has a cancer is resistant or will become resistant to a kinase inhibitor; and treating the subject with a chemotherapeutic that is not a kinase inhibitor. Also disclosed are methods of treating a subject without a kinase inhibitor resistant cancer comprising obtaining a tissue sample from a subject with a cancer, such as a kinase related cancer (e.g., ALK-related cancers); conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample, wherein the absence of a mutation in the nucleic acid sequence of a gene, region, or exon associated with kinase inhibitor resistance indicates that that the subject does not have a cancer is resistant nor will become resistant to a kinase inhibitor; and treating the subject with a kinase inhibitor.
Anaplastic Lymphoma Kinase (ALK) ALK (SEQ ID NO: 7714 (Genbank Accession No. U62540 (human coding sequence)) is a receptor tyrosine kinase (RTK) of the insulin receptor superfamily encoded by the ALK gene and is normally expressed primarily in the central and peripheral nervous systems. The 1620aa ALK polypeptide comprises a 1030aa extracellular domain which includes a 26aa amino-terminal signal peptide sequence, and binding sites located between residues 391 and 401 for the ALK ligands pleiotrophin (PTN) and midkine (MK).
Additionally, the ALK polypeptide comprises a kinase domain (residues 1116-1383) which includes three tyrosines responsible for autophosphorylation within the activation loop at residues 1278, 1282, and 1283. ALK amplification, overexpression, and mutations have been shown to constitutively activate the kinase catalytic function of the ALK
protein, with the deregulated mutant ALK in turn activating downstream cellular signaling proteins in pathways that promote aberrant cell proliferation. In fact, the mutations that result in dysregulated ALK kinase activity are associated with several types of cancers.
ALK fusions represent the most common mutation of this tyrosine kinase. Such fusions include but are not limited to nucleophosmin-ALK (NPM-ALK), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC-ALK), clathrin heavy chain-ALK (CLTC-ALK), kinesin-1 heavy chain gene-ALK (KIF5B-ALK); Ran-binding protein 2-ALK (RANBP2-ALK), SEC31L1-ALK, tropomyos in-3-ALK (TPM3-ALK), tropomyosin-4-ALK (TPM4-ALK), TRK-fused gene(Large) -ALK (TFGL-ALK), TRK-fused gene(Small) -ALK (TFGs-ALK), CARS-ALK, EML4-ALK, 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase-ALK (ATIC-ALK), AL01 7 -ALK, moesin-ALK (MSN-ALK), non-muscle myosin heavy chain gene-ALK
(MYH9-ALK), and TRK-fused gene(Extra Large) -ALK (TFGxL¨ALK). Six ALK fusions, CARS-ALK. CLTC-ALK, RANBP2-ALK, SEC31L1-ALK, TPM3-ALK, and TPM4-ALK
have been identified in IMTs. TPM3-ALK, TPM4-ALK and CLTC-ALK fusions have been detected in both classical T- or null-cell lymphomas and IMT sarcomas, whereas CARS-ALK, RANBP2-ALK, and SEC31L1-ALK occur in IMT. CLTC-ALK and NPM-ALK also occur in B-cell plasmablastic/immunoblastic lymphomas. The TPM4-ALK fusion occurs in esophageal squamous cell carcinomas, and the ALK fusion EML4-ALK, TFG-ALK and KIF5B-ALK are found in non-small cell lung cancers. EML4-ALK has also recently been identified in both colorectal and breast carcinomas as well.

ALK fusions are associated with several known cancer types. It is understood that one or more ALK fusions can be associated with a particular cancer. It is further understood that there are several types of cancer associated with ALK fusions including but not limited to anaplastic large-cell lymphoma (ALCL), neuroblastoma, breast cancer, ovarian cancer, colorectal carcinoma, non-small cell lung carcinoma, diffuse large B-cell lymphoma, esophageal squamous cell carcinoma, anaplastic large-cell lymphoma, neuroblastoma, inflammatory myofibroblastic tumors, malignant histiocytosis, and glioblastomas.
ALCL. anaplastic large-cell lymphomas comprise ¨2.5% of all NHL; within the pediatric age group specifically, ¨13% of all NHL (30 - 40% of all childhood large-cell lymphomas) are of this type. Studies of ALCL patients now divide this NHL into ALK-positive and ALK-negative subsets; ¨60% of all ALCLs are caused by ALK
fusions. For unclear reasons, ALK-positive ALCL patients fare significantly better following CHOP
based multi-agent conventional chemotherapy than those with ALK-negative disease (with overall 5-year survivals of ¨75% vs. ¨35%, respectively). However, more than a third of patients suffer multiple relapses following chemotherapy, thus the 5-year disease-free survival of ALK-positive ALCL is only ¨40%.
ALK+ Diffuse large B-cell lymphoma. In 2003, ALK fusions were shown to occur in a non-ALCL form of NHL with the description of CLTC-ALK or NPM-ALK in diffuse large B-cell lymphomas (ALK+ DLBCLs). Consistent with their B-lineage, these NHLs express cytoplasmic IgA and plasma cell markers, and possess an immunoblastic morphology. Translational research studies revealed the t(2;17) and CLTC-ALK
mRNA in the majority of these lymphomas, while immunolabeling confirmed granular ALK
staining identical to that observed in CLTC-ALK-positive ALCL. As for all other ALK
fusion partner proteins, a self-association motif in the CLTC portion of CLTC-ALK
mediates constitutive self-association and activation of the fusion kinase to drive lymphomagenesis.
ALK+ DLBCLs occur predominately in adults; however, the t(2;5) and NPM-ALK
mRNA
in pediatric lymphomas are phenotypically identical to CLTC-ALK- positive adult B-NHLs.

Approximately 0.5-1% of all DLBCL is thought to be ALK-positive. The identification of DLBCLs caused by mutant ALK is important because patients with these lymphomas have outcomes that are much inferior to ALK-negative DLBCL patients following CHOP-based treatments; thus, ALK+ DLBCL patients should strongly be considered as candidates for ALK-targeted kinase inhibitor therapy.
ALK+ systemic histiocytosis. ALK fusions were described in 2008 in another hematopoietic neoplasm, systemic histiocytosis. Three cases of this previously uncharacterized form of histiocytosis, which presents in early infancy, exhibited ALK
immunoreactivity and the one case analyzed molecularly expressed TPM3-ALK.
In addition to the aforementioned hematological malignancies in which constitutively activated ALK fusions have been shown to be a causative mechanism in many cases, the genesis of subsets of various solid tumors in some instances, very common human tumors such as non-small cell lung cancer, colorectal and breast cancers has recently been demonstrated to be due to aberrantly activated ALK.
Inflammatory myofibroblastic tumor. The first non-hematopoietic tumor discovered to express ALK fusions was the sarcoma known as inflammatory myofibroblastic tumor (IMT), a spindle cell proliferation in the soft tissue and viscera of children and young adults (mean age at diagnosis ¨10 years). Many IMTs are indolent and can be cured by resection.
However, locally recurrent, invasive, and metastatic IMTs are not uncommon and current chemo- and radio-therapies are completely ineffective. Disclosed herein is the involvement of chromosome 2p23 (the location of the ALK gene) in IMTs, as well as ALK gene rearrangement. ALK immunoreactivity in 7 of 11 IMTs has been shown and TPM3-ALK
and TPM4-ALK were identified in several cases. Additionally, two additional ALK fusions in IMT, CLTC- and RanBP2-ALK were identified. ALK fusions have also been examined by immunostaining in 73 IMTs, finding 60% (44 of the 73 cases) to be ALK-positive. Thus, ALK deregulation is of pathogenic importance in a majority of IMTs.
Non-small cell lung carcinoma. The role of ALK fusions in cancer expanded further with the description of the novel EML4-ALK chimeric protein in 5 of 75 (6.7%) Japanese non-small cell lung carcinoma patients. Shortly thereafter, the existence of ALK fusions in lung cancer was corroborated by a different group who found 6 of 137 (4.4%) Chinese lung cancer patients to express ALK fusions (EML4-ALK, 3 pts; TFG-ALK, 1 pt; X-ALK.
Two common themes have emerged ¨ 1) ALK fusions occur predominately in patients with adenocarcinoma (although occasional ALK-positive NSCLCs of squamous or mixed histologies are observed), mostly in individuals with minimal/no smoking history, and 2) ALK abnormalities usually occur exclusive of other common genetic abnormalities (e.g., EGFR and KRAS mutations). The exact percentage of NSCLCs caused by ALK fusions is not yet clear but estimates based on reports in the biomedical literature suggest a range of ¨5-10%.
Esophageal squamous cell carcinoma. In 45 Iranian patients, a proteomics approach identified proteins under or over-represented in esophageal squamous cell carcinomas (ESCCs); TPM4-ALK was among those proteins over-represented. A second proteomics-based ESCC study ¨ in this case, in Chinese patients ¨ identified TPM4-ALK in these tumors as well.
Colorectal carcinoma, breast cancer. Three human tumor types ¨ colorectal, breast, and non-small cell lung cancers were surveyed for the presence of the EML4-ALK
fusion (other ALK mutations were not assessed in this study). In addition to confirming the expression of EML4-ALK in NSCLC (in 12 of 106 specimens studied, 11.3%), a subsets of breast (5 of 209 cases, 2.4%) and colorectal (2 of 83 cases, 2.4%) carcinomas were EML4-ALK-positive. In addition to known EML4-ALK variants 1 (E13; A20) and 2 (E20;
A20), a novel variant (E21; A20) was found in colorectal carcinoma.
ALK in familial and sporadic neuroblastoma. Neuroblastoma is the most common extracranial solid tumor of childhood, and is derived from the developing neural crest. A
small subset (-1-2%) of neuroblastomas exhibit a familial predisposition with an autosomal dominant inheritance. Most neuroblastoma patients have aggressive disease associated with survival probabilities <40% despite intensive chemo- and radio-therapy, and the disease accounts for ¨15% of all childhood cancer mortality. ALK had previously been found to be constitutively activated also due to high-level over-expression as a result of gene amplification in a small number of neuroblastoma cell lines, in fact, ALK
amplification occurs in ¨15% of neuroblastomas in addition to activating point mutations.
These missense mutations in ALK have been confirmed as activating mutations that drive neuroblastoma growth; furthermore, incubation of neuroblastoma cell lines with ALK small-molecule inhibitors reveal those cells with ALK activation (but not cell lines with normal levels of expression of wild-type ALK) to exhibit robust cytotoxic responses.
The sensitive detection of a mutation at a known site in DNA is readily done with existing technologies. Allele specific primers can be designed to target a mutation at a known location such that its signal can be preferentially amplified over wild-type DNA.
Next Generation Sequencing for Genetic Testing From a technical perspective High-throughput or Next Generation Sequencing (NGS) represents an attractive option for detecting the somatic mutations within a gene.
Unlike PCR, microarrays, high-resolution melting and mass spectrometry, which all indirectly infer sequence content, NGS directly ascertains the identity of each base and the order in which they fall within a gene. The newest platforms on the market have the capacity to cover an exonic region 10,000 times over, meaning the content of each base position in the sequence is measured thousands of different times. This high level of coverage ensures that the consensus sequence is extremely accurate and enables the detection of rare variants within a heterogeneous sample. For example, in a sample extracted from FFPE tissue, relevant mutations are only present at a frequency of 1% with the wild-type allele comprising the remainder. When this sample is sequenced at 10,000X
coverage, then even the rare allele, comprising only 1% of the sample, is uniquely measured 100 times over. Thus, NGS can provide reliably accurate results with very high sensitivity, making it ideal for clinical diagnostic testing of FFPEs and other mixed samples.
In one aspect, disclosed herein are methods and assays for detecting kinase inhibitor resistance or determining the susceptibility to a particular kinase inhibitor treatment in an ALK-related cancer comprising performing next generation sequencing on a tissue sample obtained from a subject with an ALK-related cancer, wherein the presence of a nucleic acid variation in the ALK, BRAF, EGFR, KIT, or KRAS sequence of the tissue sample at a nucleic acid residue indicates that presence of kinase inhibitor resistance.
For example, the methods and assays for detecting kinase inhibitor resistance or determining the susceptibility or developing kinase inhibitor resistance in an ALK-related cancer or determining the suitability of a particular kinase inhibitor for use in treating an ALK-related cancer in a subject can comprise the detection of any of the mutations in Tables 2-6. It is understood that the methods and assays can further comprise comparing the sequence to known kinase inhibitor resistance mutations list and determining what if any kinase inhibitors are affected by the mutation and altering or maintaining treatment as appropriate to utilize kinase inhibitors that are unaffected by the mutation. As the disclosed methods and assays employ the use of primers or primer sets to detect mutations that confer kinase inhibitor resistance, also disclosed herein are primer panels for use in next generation sequencing for the determination of kinase inhibitor resistance comprising one or more primer sets from each of KIT, BRAF, KRAS, EGFR, and ALK, for example, the disclosed primer panels, methods, and assays can comprise one or more of the primers or primer sets listed in Tables 7-14.
Examples of Next Generation Sequencing techniques include, but are not limited to Massively Parallel Signature Sequencing (MPSS), Polony sequencing, pyrosequencing, Reversible dye-terminator sequencing, SOLiD sequencing, Ion semiconductor sequencing, DNA nanoball sequencing, Helioscope single molecule sequencing, Single molecule real time (SMRT) sequencing, Single molecule real time (RNAP) sequencing, and Nanopore DNA sequencing.
MPSS was a bead-based method that used a complex approach of adapter ligation followed by adapter decoding, reading the sequence in increments of four nucleotides; this method made it susceptible to sequence-specific bias or loss of specific sequences.

Polony sequencing, combined an in vitro paired-tag library with emulsion PCR, an automated microscope, and ligation-based sequencing chemistry to sequence an E. coli genome at an accuracy of > 99.9999% and a cost approximately 1/10 that of Sanger sequencing.
A parallelized version of pyrosequencing, the method amplifies DNA inside water droplets in an oil solution (emulsion PCR), with each droplet containing a single DNA
template attached to a single primer-coated bead that then forms a clonal colony. The sequencing machine contains many picolitre-volume wells each containing a single bead and sequencing enzymes. Pyrosequencing uses luciferase to generate light for detection of the individual nucleotides added to the nascent DNA, and the combined data are used to generate sequence read-outs. This technology provides intermediate read length and price per base compared to Sanger sequencing on one end and Solexa and SOLiD on the other.
A sequencing technology based on reversible dye-terminators. DNA molecules are first attached to primers on a slide and amplified so that local clonal colonies are formed.
Four types of reversible terminator bases (RT-bases) are added, and non-incorporated nucleotides are washed away. Unlike pyrosequencing, the DNA can only be extended one nucleotide at a time. A camera takes images of the fluorescently labeled nucleotides, then the dye along with the terminal 3' blocker is chemically removed from the DNA, allowing the next cycle.
SOLiD technology employs sequencing by ligation. Here, a pool of all possible oligonucleotides of a fixed length are labeled according to the sequenced position.
Oligonucleotides are annealed and ligated; the preferential ligation by DNA
ligase for matching sequences results in a signal informative of the nucleotide at that position. Before sequencing, the DNA is amplified by emulsion PCR. The resulting bead, each containing only copies of the same DNA molecule, are deposited on a glass slide. The result is sequences of quantities and lengths comparable to Illumina sequencing.
Ion semiconductor sequencing is based on using standard sequencing chemistry, but with a novel, semiconductor based detection system. This method of sequencing is based on the detection of hydrogen ions that are released during the polymerization of DNA, as opposed to the optical methods used in other sequencing systems. A microwell containing a template DNA strand to be sequenced is flooded with a single type of nucleotide. If the introduced nucleotide is complementary to the leading template nucleotide it is incorporated into the growing complementary strand. This causes the release of a hydrogen ion that triggers a hypersensitive ion sensor, which indicates that a reaction has occurred. If homopolymer repeats are present in the template sequence multiple nucleotides will be incorporated in a single cycle. This leads to a corresponding number of released hydrogens and a proportionally higher electronic signal.
DNA nanoball sequencing is a type of high throughput sequencing technology used to determine the entire genomic sequence of an organism. The method uses rolling circle replication to amplify small fragments of genomic DNA into DNA nanoballs.
Unchained sequencing by ligation is then used to determine the nucleotide sequence. This method of DNA sequencing allows large numbers of DNA nanoballs to be sequenced per run.
Helicos's single-molecule sequencing uses DNA fragments with added polyA tail adapters, which are attached to the flow cell surface. The next steps involve extension-based sequencing with cyclic washes of the flow cell with fluorescently labeled nucleotides (one nucleotide type at a time, as with the Sanger method). The reads are performed by the Helioscope sequencer.
SMRT sequencing is based on the sequencing by synthesis approach. The DNA is synthesized in zero-mode wave-guides (ZMWs) - small well-like containers with the capturing tools located at the bottom of the well. The sequencing is performed with use of unmodified polymerase (attached to the ZMW bottom) and fluorescently labeled nucleotides flowing freely in the solution. The wells are constructed in a way that only the fluorescence occurring by the bottom of the well is detected. The fluorescent label is detached from the nucleotide at its incorporation into the DNA strand, leaving an unmodified DNA strand.

Single molecule real time sequencing based on RNA polymerase (RNAP), which is attached to a polystyrene bead, with distal end of sequenced DNA is attached to another bead, with both beads being placed in optical traps. RNAP motion during transcription brings the beads in closer and their relative distance changes, which can then be recorded at a single nucleotide resolution. The sequence is deduced based on the four readouts with lowered concentrations of each of the four nucleotide types (similarly to Sangers method).
Nanopore sequencing is based on the readout of electrical signal occurring at nucleotides passing by alpha-hemolysin pores covalently bound with cyclodextrin. The DNA passing through the nanopore changes its ion current. This change is dependent on the shape, size and length of the DNA sequence. Each type of the nucleotide blocks the ion flow through the pore for a different period of time.
VisiGen Biotechnologies uses a specially engineered DNA polymerase. This polymerase acts as a sensor - having incorporated a donor fluorescent dye by its active centre. This donor dye acts by FRET (fluorescent resonant energy transfer), inducing fluorescence of differently labeled nucleotides. This approach allows reads performed at the speed at which polymerase incorporates nucleotides into the sequence (several hundred per second). The nucleotide fluorochrome is released after the incorporation into the DNA
strand.
Sequencing by hybridization is a non-enzymatic method that uses a DNA
microan-ay. A single pool of DNA whose sequence is to be determined is fluorescently labeled and hybridized to an array containing known sequences. Strong hybridization signals from a given spot on the array identify its sequence in the DNA being sequenced.
Mass spectrometry may be used to determine mass differences between DNA
fragments produced in chain-termination reactions.
Another NGS approach is sequencing by synthesis (SBS) technology which is capable of overcoming the limitations of existing pyrosequencing based NGS
platforms.
Such technologies rely on complex enzymatic cascades for read out, are unreliable for the accurate determination of the number of nucleotides in homopolymeric regions and require excessive amounts of time to run individual nucleotides across growing DNA
strands. The SBS NGS platform uses a direct sequencing approach to produce a sequencing strategy with very a high precision, rapid pace and low cost.
SBS sequencing is initialized by fragmenting of the template DNA into fragments, amplification, annealing of DNA sequencing primers, and finally affixing as a high-density array of spots onto a glass chip. The array of DNA fragments are sequenced by extending each fragment with modified nucleotides containing cleavable chemical moieties linked to fluorescent dyes capable of discriminating all four possible nucleotides. The array is scanned continuously by a high-resolution electronic camera (Measure) to determine the fluorescent intensity of each base (A, C, G or T) that was newly incorporated into the extended DNA fragment. After the incorporation of each modified base the array is exposed to cleavage chemistry to break off the fluorescent dye and end cap allowing additional bases to be added. The process is then repeated until the fragment is completely sequenced or maximal read length has been achieved.
mRNA detection and quantification A number of widely used procedures exist for detecting and determining the abundance of a particular mRNA in a total or poly(A) RNA sample. For example, specific mRNAs can be detected using Northern blot analysis, nuclease protection assays (NPA), in situ hybridization (e.g., fluorescence in situ hybridization (FISH)), or reverse transcription-polymerase chain reaction (RT-PCR), and microarray.
In theory, each of these techniques can be used to detect specific RNAs and to precisely determine their expression level. In general, Northern analysis is the only method that provides information about transcript size, whereas NPAs are the easiest way to simultaneously examine multiple messages. In situ hybridization is used to localize expression of a particular gene within a tissue or cell type, and RT-PCR is the most sensitive method for detecting and quantitating gene expression.
RT-PCR allows for the detection of the RNA transcript of any gene, regardless of the scarcity of the starting material or relative abundance of the specific mRNA. In RT-PCR, an RNA template is copied into a complementary DNA (cDNA) using a retroviral reverse transcriptase. The cDNA is then amplified exponentially by PCR using a DNA
polymerase. The reverse transcription and PCR reactions can occur in the same or difference tubes. RT-PCR is somewhat tolerant of degraded RNA. As long as the RNA is intact within the region spanned by the primers, the target will be amplified.
Relative quantitative RT-PCR involves amplifying an internal control simultaneously with the gene of interest. The internal control is used to normalize the samples. Once normalized, direct comparisons of relative abundance of a specific mRNA
can be made across the samples. It is crucial to choose an internal control with a constant level of expression across all experimental samples (i.e., not affected by experimental treatment). Commonly used internal controls (e.g., GAPDH, 13-actin, cyclophilin) often vary in expression and, therefore, may not be appropriate internal controls.
Additionally, most common internal controls are expressed at much higher levels than the mRNA
being studied. For relative RT-PCR results to be meaningful, all products of the PCR
reaction must be analyzed in the linear range of amplification. This becomes difficult for transcripts of widely different levels of abundance.
Competitive RT-PCR is used for absolute quantitation. This technique involves designing, synthesizing, and accurately quantitating a competitor RNA that can be distinguished from the endogenous target by a small difference in size or sequence. Known amounts of the competitor RNA are added to experimental samples and RT-PCR is performed. Signals from the endogenous target are compared with signals from the competitor to determine the amount of target present in the sample.
Northern analysis is the easiest method for determining transcript size, and for identifying alternatively spliced transcripts and multigene family members. It can also be used to directly compare the relative abundance of a given message between all the samples on a blot. The Northern blotting procedure is straightforward and provides opportunities to evaluate progress at various points (e.g., intactness of the RNA sample and how efficiently it has transferred to the membrane). RNA samples are first separated by size via electrophoresis in an agarose gel under denaturing conditions. The RNA is then transferred to a membrane, crosslinked and hybridized with a labeled probe. Nonisotopic or high specific activity radiolabeled probes can be used including random-primed, nick-translated, or PCR-generated DNA probes, in vitro transcribed RNA probes, and oligonucleotides.
Additionally, sequences with only partial homology (e.g., cDNA from a different species or genomic DNA fragments that might contain an exon) may be used as probes.
The Nuclease Protection Assay (NPA) (including both ribonuclease protection assays and Si nuclease assays) is a sensitive method for the detection and quantitation of specific mRNAs. The basis of the NPA is solution hybridization of an antisense probe (radiolabeled or nonisotopic) to an RNA sample. After hybridization, single-stranded, unhybridized probe and RNA are degraded by nucleases. The remaining protected fragments are separated on an acrylamide gel. Solution hybridization is typically more efficient than membrane-based hybridization, and it can accommodate up to 100 lug of sample RNA, compared with the 20-30 1..tg maximum of blot hybridizations. NPAs are also less sensitive to RNA sample degradation than Northern analysis since cleavage is only detected in the region of overlap with the probe (probes are usually about 100-400 bases in length).
NPAs are the method of choice for the simultaneous detection of several RNA
species. During solution hybridization and subsequent analysis, individual probe/target interactions are completely independent of one another. Thus, several RNA
targets and appropriate controls can be assayed simultaneously (up to twelve have been used in the same reaction), provided that the individual probes are of different lengths.
NPAs are also commonly used to precisely map mRNA termini and intron/exon junctions.
In situ hybridization (ISH) is a powerful and versatile tool for the localization of specific mRNAs in cells or tissues. Unlike Northern analysis and nuclease protection assays, ISH does not require the isolation or electrophoretic separation of RNA.
Hybridization of the probe takes place within the cell or tissue. Since cellular structure is maintained throughout the procedure, ISH provides information about the location of mRNA within the tissue sample.
The procedure begins by fixing samples in neutral-buffered formalin, and embedding the tissue in paraffin. The samples are then sliced into thin sections and mounted onto microscope slides. (Alternatively, tissue can be sectioned frozen and post-fixed in paraformaldehyde.) After a series of washes to dewax and rehydrate the sections, a Proteinase K digestion is performed to increase probe accessibility, and a labeled probe is then hybridized to the sample sections. Radiolabeled probes are visualized with liquid film dried onto the slides, while non-isotopically labeled probes are conveniently detected with colorimetric or fluorescent reagents.
DNA detection and quantification The methods, assays, and primer panels disclosed herein relate to the detection of nucleic acid variation that confer kinase inhibitor resistance in the form of, for example, point mutations and truncations of, KRAS, BRAF, KIT, EGFR, and/or ALK. Thus, in one aspect, disclosed herein are methods, assays, and use of the disclosed primer panels for diagnosing an anaplastic lymphoma kinase (ALK) related cancer in a subject is resistant to a kinase inhibitor comprise performing NGS which sequences DNA from a tissue sample from the subject. It is understood that high throughput sequencing methods (also known as next generation sequencing methods) can comprise any known amplification and detection method for DNA known in the art.
A number of widely used procedures exist for detecting and determining the abundance of a particular DNA in a sample. For example, the technology of PCR
permits amplification and subsequent detection of minute quantities of a target nucleic acid. Details of PCR are well described in the art, including, for example, U.S. Pat. Nos.
4,683,195 to Mullis et al., 4,683,202 to Mullis and 4,965,188 to Mullis et al. Generally, oligonucleotide primers are annealed to the denatured strands of a target nucleic acid, and primer extension products are formed by the polymerization of deoxynucleoside triphosphates by a polymerase. A typical PCR method involves repetitive cycles of template nucleic acid denaturation, primer annealing and extension of the annealed primers by the action of a thermostable polymerase. The process results in exponential amplification of the target nucleic acid, and thus allows the detection of targets existing in very low concentrations in a sample. It is understood and herein contemplated that there are variant PCR
methods known in the art that may also be utilized in the disclosed methods, for example, Quantitative PCR (QPCR); microarrays, real-time PCR; hot start PCR; nested PCR; allele-specific PCR; and Touchdown PCR.
Microarrays An array is an orderly arrangement of samples, providing a medium for matching known and unknown DNA samples based on base-pairing rules and automating the process of identifying the unknowns. An array experiment can make use of common assay systems such as microplates or standard blotting membranes, and can be created by hand or make use of robotics to deposit the sample. In general, arrays are described as macroarrays or microarrays, the difference being the size of the sample spots. Macroarrays contain sample spot sizes of about 300 microns or larger and can be easily imaged by existing gel and blot scanners. The sample spot sizes in microarray can be 300 microns or less, but typically less than 200 microns in diameter and these arrays usually contains\ thousands of spots.
Microarrays require specialized robotics and/or imaging equipment that generally are not commercially available as a complete system. Terminologies that have been used in the literature to describe this technology include, but not limited to: biochip, DNA chip, DNA
microarray, GENECHIPO (Affymetrix, Inc which refers to its high density, oligonucleotide-based DNA arrays), and gene array.
DNA microarrays, or DNA chips are fabricated by high-speed robotics, generally on glass or nylon substrates, for which probes with known identity are used to determine complementary binding, thus allowing massively parallel gene expression and gene discovery studies. An experiment with a single DNA chip can provide information on thousands of genes simultaneously. It is herein contemplated that the disclosed microarrays can be used to monitor gene expression, disease diagnosis, gene discovery, drug discovery (pharmacogenomics), and toxicological research or toxicogenomics.
There are two variants of the DNA microarray technology, in terms of the property of arrayed DNA sequence with known identity. Type I microarrays comprise a probe cDNA (500-5,000 bases long) that is immobilized to a solid surface such as glass using robot spotting and exposed to a set of targets either separately or in a mixture. This method is traditionally referred to as DNA microarray. With Type I microarrays, localized multiple copies of one or more polynucleotide sequences, preferably copies of a single polynucleotide sequence are immobilized on a plurality of defined regions of the substrate's surface. A polynucleotide refers to a chain of nucleotides ranging from 5 to

10,000 nucleotides. These immobilized copies of a polynucleotide sequence are suitable for use as probes in hybridization experiments.
To prepare beads coated with immobilized probes, beads are immersed in a solution containing the desired probe sequence and then immobilized on the beads by covalent or non-covalent means. Alternatively, when the probes are immobilized on rods, a given probe can be spotted at defined regions of the rod. Typical dispensers include a micropipette delivering solution to the substrate with a robotic system to control the position of the micropipette with respect to the substrate. There can be a multiplicity of dispensers so that reagents can be delivered to the reaction regions simultaneously. In one embodiment, a microarray is formed by using ink-jet technology based on the piezoelectric effect, whereby a narrow tube containing a liquid of interest, such as oligonucleotide synthesis reagents, is encircled by an adapter. An electric charge sent across the adapter causes the adapter to expand at a different rate than the tube and forces a small drop of liquid onto a substrate.
Samples may be any sample containing polynucleotides (polynucleotide targets) of interest and obtained from any bodily fluid (blood, urine, saliva, phlegm, gastric juices, etc.), cultured cells, biopsies, or other tissue preparations. DNA or RNA can be isolated from the sample according to any of a number of methods well known to those of skill in the art. In one embodiment, total RNA is isolated using the TRIzol total RNA
isolation reagent (Life Technologies, Inc., Rockville, Md.) and RNA is isolated using oligo d(T) column chromatography or glass beads. After hybridization and processing, the hybridization signals obtained should reflect accurately the amounts of control target polynucleotide added to the sample.
The plurality of defined regions on the substrate can be arranged in a variety of formats. For example, the regions may be arranged perpendicular or in parallel to the length of the casing. Furthermore, the targets do not have to be directly bound to the substrate, but rather can be bound to the substrate through a linker group. The linker groups may typically vary from about 6 to 50 atoms long. Linker groups include ethylene glycol oligomers, diamines, diacids and the like. Reactive groups on the substrate surface react with one of the terminal portions of the linker to bind the linker to the substrate. The other terminal portion of the linker is then functionalized for binding the probes.
Sample polynucleotides may be labeled with one or more labeling moieties to allow for detection of hybridized probe/target polynucleotide complexes. The labeling moieties can include compositions that can be detected by spectroscopic, photochemical, biochemical, bioelectronic, immunochemical, electrical, optical or chemical means. The labeling moieties include radioisotopes, such as 32P, 33P or 35, chemiluminescent compounds, labeled binding proteins, heavy metal atoms, spectroscopic markers, such as fluorescent markers and dyes, magnetic labels, linked enzymes, mass spectrometry tags, spin labels, electron transfer donors and acceptors, biotin, and the like.
Labeling can be carried out during an amplification reaction, such as polymerase chain reaction and in vitro or in vivo transcription reactions. Alternatively, the labeling moiety can be incorporated after hybridization once a probe-target complex his formed. In one embodiment, biotin is first incorporated during an amplification step as described above. After the hybridization reaction, unbound nucleic acids are rinsed away so that the only biotin remaining bound to the substrate is that attached to target polynucleotides that are hybridized to the polynucleotide probes. Then, an ayidin-conjugated fluorophore, such as ayidin-phycoerythrin, that binds with high affinity to biotin is added.

Hybridization causes a polynucleotide probe and a complementary target to form a stable duplex through base pairing. Hybridization methods are well known to those skilled in the art. Stringent conditions for hybridization can be defined by salt concentration, temperature, and other chemicals and conditions. Varying additional parameters, such as hybridization time, the concentration of detergent (sodium dodecyl sulfate, SDS) or solvent (formamide), and the inclusion or exclusion of carrier DNA, are well known to those skilled in the art. Additional variations on these conditions will be readily apparent to those skilled in the art.
Methods for detecting complex formation are well known to those skilled in the art.
In one embodiment, the polynucleotide probes are labeled with a fluorescent label and measurement of levels and patterns of complex formation is accomplished by fluorescence microscopy, preferably confocal fluorescence microscopy. An argon ion laser excites the fluorescent label, emissions are directed to a photomultiplier and the amount of emitted light detected and quantitated. The detected signal should be proportional to the amount of probe/target polynucleotide complex at each position of the microarray. The fluorescence microscope can be associated with a computer-driven scanner device to generate a quantitative two-dimensional image of hybridization intensities. The scanned image is examined to determine the abundance/expression level of each hybridized target polynucleotide.
In a differential hybridization experiment, polynucleotide targets from two or more different biological samples are labeled with two or more different fluorescent labels with different emission wavelengths. Fluorescent signals are detected separately with different photomultipliers set to detect specific wavelengths. The relative abundances/expression levels of the target polynucleotides in two or more samples is obtained.
Typically, microarray fluorescence intensities can be normalized to take into account variations in hybridization intensities when more than one microarray is used under similar test conditions. In one embodiment, individual polynucleotide probe/target complex hybridization intensities are normalized using the intensities derived from internal normalization controls contained on each microarray.
Type II microarrays comprise an array of oligonucleotides (20-80-mer oligos) or peptide nucleic acid (PNA) probes that is synthesized either in situ (on-chip) or by conventional synthesis followed by on-chip immobilization. The array is exposed to labeled sample DNA, hybridized, and the identity/abundance of complementary sequences are determined. This method, "historically" called DNA chips, was developed at Affymetrix, Inc. , which sells its photolithographically fabricated products under the GENECHIPO
trademark.
The basic concept behind the use of Type II arrays for gene expression is simple:
labeled cDNA or cRNA targets derived from the mRNA of an experimental sample are hybridized to nucleic acid probes attached to the solid support. By monitoring the amount of label associated with each DNA location, it is possible to infer the abundance of each mRNA species represented. Although hybridization has been used for decades to detect and quantify nucleic acids, the combination of the miniaturization of the technology and the large and growing amounts of sequence information, have enormously expanded the scale at which gene expression can be studied.
Microarray manufacturing can begin with a 5-inch square quartz wafer.
Initially the quartz is washed to ensure uniform hydroxylation across its surface. Because quartz is naturally hydroxylated, it provides an excellent substrate for the attachment of chemicals, such as linker molecules, that are later used to position the probes on the arrays.
The wafer is placed in a bath of silane, which reacts with the hydroxyl groups of the quartz, and forms a matrix of covalently linked molecules. The distance between these silane molecules determines the probes' packing density, allowing arrays to hold over 500,000 probe locations, or features, within a mere 1.28 square centimeters.
Each of these features harbors millions of identical DNA molecules. The silane film provides a uniform hydroxyl density to initiate probe assembly. Linker molecules, attached to the silane matrix, provide a surface that may be spatially activated by light.

Probe synthesis occurs in parallel, resulting in the addition of an A, C, T, or G
nucleotide to multiple growing chains simultaneously. To define which oligonucleotide chains will receive a nucleotide in each step, photolithographic masks, carrying 18 to 20 square micron windows that correspond to the dimensions of individual features, are placed over the coated wafer. The windows are distributed over the mask based on the desired sequence of each probe. When ultraviolet light is shone over the mask in the first step of synthesis, the exposed linkers become deprotected and are available for nucleotide coupling.
Once the desired features have been activated, a solution containing a single type of deoxynucleotide with a removable protection group is flushed over the wafer's surface. The nucleotide attaches to the activated linkers, initiating the synthesis process.
Although each position in the sequence of an oligonucleotide can be occupied by lof 4nucleotides, resulting in an apparent need for 25 x 4, or 100, different masks per wafer, the synthesis process can be designed to significantly reduce this requirement.
Algorithms that help minimize mask usage calculate how to best coordinate probe growth by adjusting synthesis rates of individual probes and identifying situations when the same mask can be used multiple times.
Some of the key elements of selection and design are common to the production of all microarrays, regardless of their intended application. Strategies to optimize probe hybridization, for example, are invariably included in the process of probe selection.
Hybridization under particular pH, salt, and temperature conditions can be optimized by taking into account melting temperatures and using empirical rules that correlate with desired hybridization behaviors.
To obtain a complete picture of a gene's activity, some probes are selected from regions shared by multiple splice or polyadenylation variants. In other cases, unique probes that distinguish between variants are favored. Inter-probe distance is also factored into the selection process.
A different set of strategies is used to select probes for genotyping arrays that rely on multiple probes to interrogate individual nucleotides in a sequence. The identity of a target base can be deduced using four identical probes that vary only in the target position, each containing one of the four possible bases.
Alternatively, the presence of a consensus sequence can be tested using one or two probes representing specific alleles. To genotype heterozygous or genetically mixed samples, arrays with many probes can be created to provide redundant information, resulting in unequivocal genotyping. In addition, generic probes can be used in some applications to maximize flexibility. Some probe arrays, for example, allow the separation and analysis of individual reaction products from complex mixtures, such as those used in some protocols to identify single nucleotide polymorphisms (SNPs).
Real-time PCR
Real-time PCR monitors the fluorescence emitted during the reaction as an indicator of amplicon production during each PCR cycle (i.e., in real time) as opposed to the endpoint detection. The real-time progress of the reaction can be viewed in some systems. Real-time PCR does not detect the size of the amplicon and thus does not allow the differentiation between DNA and cDNA amplification, however, it is not influenced by non-specific amplification unless SYBR Green is used. Real-time PCR quantitation eliminates post-PCR
processing of PCR products. This helps to increase throughput and reduce the chances of carryover contamination. Real-time PCR also offers a wide dynamic range of up to 107-fold.
Dynamic range of any assay determines how much target concentration can vary and still be quantified. A wide dynamic range means that a wide range of ratios of target and normaliser can be assayed with equal sensitivity and specificity. It follows that the broader the dynamic range, the more accurate the quantitation. When combined with RT-PCR, a real-time RT-PCR reaction reduces the time needed for measuring the amount of amplicon by providing for the visualization of the amplicon as the amplification process is progressing.
The real-time PCR system is based on the detection and quantitation of a fluorescent reporter. This signal increases in direct proportion to the amount of PCR
product in a reaction. By recording the amount of fluorescence emission at each cycle, it is possible to monitor the PCR reaction during exponential phase where the first significant increase in the amount of PCR product correlates to the initial amount of target template.
The higher the starting copy number of the nucleic acid target, the sooner a significant increase in fluorescence is observed. A significant increase in fluorescence above the baseline value measured during the 3-15 cycles can indicate the detection of accumulated PCR
product.
A fixed fluorescence threshold is set significantly above the baseline that can be altered by the operator. The parameter CT (threshold cycle) is defined as the cycle number at which the fluorescence emission exceeds the fixed threshold.
There are three main fluorescence-monitoring systems for DNA amplification:
(1) hydrolysis probes; (2) hybridising probes; and (3) DNA-binding agents.
Hydrolysis probes include TaqMan probes, molecular beacons and scorpions. They use the fluorogenic 5' exonuclease activity of Taq polymerase to measure the amount of target sequences in cDNA
samples.
TaqMan probes are oligonucleotides longer than the primers (20-30 bases long with a Tm value of 10 C higher) that contain a fluorescent dye usually on the 5' base, and a quenching dye (usually TAMRA) typically on the 3' base. When irradiated, the excited fluorescent dye transfers energy to the nearby quenching dye molecule rather than fluorescing (this is called FRET = Forster or fluorescence resonance energy transfer). Thus, the close proximity of the reporter and quencher prevents emission of any fluorescence while the probe is intact. TaqMan probes are designed to anneal to an internal region of a PCR product. When the polymerase replicates a template on which a TaqMan probe is bound, its 5' exonuclease activity cleaves the probe. This ends the activity of quencher (no FRET) and the reporter dye starts to emit fluorescence which increases in each cycle proportional to the rate of probe cleavage. Accumulation of PCR products is detected by monitoring the increase in fluorescence of the reporter dye (note that primers are not labelled). TaqMan assay uses universal thermal cycling parameters and PCR
reaction conditions. Because the cleavage occurs only if the probe hybridises to the target, the origin of the detected fluorescence is specific amplification. The process of hybridisation and cleavage does not interfere with the exponential accumulation of the product.
One specific requirement for fluorogenic probes is that there be no G at the 5' end. A 'G' adjacent to the reporter dye can quench reporter fluorescence even after cleavage.
Molecular beacons also contain fluorescent (FAM, TAMRA, TET, ROX) and quenching dyes (typically DABCYL) at either end but they are designed to adopt a hairpin structure while free in solution to bring the fluorescent dye and the quencher in close proximity for FRET to occur. They have two arms with complementary sequences that form a very stable hybrid or stem. The close proximity of the reporter and the quencher in this hairpin configuration suppresses reporter fluorescence. When the beacon hybridises to the target during the annealing step, the reporter dye is separated from the quencher and the reporter fluoresces (FRET does not occur). Molecular beacons remain intact during PCR
and must rebind to target every cycle for fluorescence emission. This will correlate to the amount of PCR product available. All real-time PCR chemistries allow detection of multiple DNA species (multiplexing) by designing each probe/beacon with a spectrally unique fluor/quench pair as long as the platform is suitable for melting curve analysis if SYBR
green is used. By multiplexing, the target(s) and endogenous control can be amplified in single tube.
With Scorpion probes, sequence-specific priming and PCR product detection is achieved using a single oligonucleotide. The Scorpion probe maintains a stem-loop configuration in the unhybridised state. The fluorophore is attached to the 5' end and is quenched by a moiety coupled to the 3' end. The 3' portion of the stem also contains sequence that is complementary to the extension product of the primer. This sequence is linked to the 5' end of a specific primer via a non-amplifiable monomer. After extension of the Scorpion primer, the specific probe sequence is able to bind to its complement within the extended amplicon thus opening up the hairpin loop. This prevents the fluorescence from being quenched and a signal is observed.
Another alternative is the double-stranded DNA binding dye chemistry, which quantitates the amplicon production (including non-specific amplification and primer-dimer complex) by the use of a non-sequence specific fluorescent intercalating agent (SYBR-green I or ethidium bromide). It does not bind to ssDNA. SYBR green is a fluorogenic minor groove binding dye that exhibits little fluorescence when in solution but emits a strong fluorescent signal upon binding to double-stranded DNA. Disadvantages of SYBR
green-based real-time PCR include the requirement for extensive optimisation.
Furthermore, non-specific amplifications require follow-up assays (melting point curve or dissociation analysis) for amplicon identification. The method has been used in HFE-C282Y
genotyping.
Another controllable problem is that longer amplicons create a stronger signal (if combined with other factors, this may cause CDC camera saturation, see below). Normally SYBR
green is used in singleplex reactions, however when coupled with melting point analysis, it can be used for multiplex reactions.
The threshold cycle or the CT value is the cycle at which a significant increase in ARn is first detected (for definition of ARn, see below). The threshold cycle is when the system begins to detect the increase in the signal associated with an exponential growth of PCR product during the log-linear phase. This phase provides the most useful information about the reaction (certainly more important than the end-point). The slope of the log-linear phase is a reflection of the amplification efficiency. The efficiency (Eff) of the reaction can be calculated by the formula: Eff=10(-1/s10Pe)-1. The efficiency of the PCR
should be 90 -100% (3.6> slope >3.1). A number of variables can affect the efficiency of the PCR. These factors include length of the amplicon, secondary structure and primer quality. Although valid data can be obtained that fall outside of the efficiency range, the qRT-PCR should be further optimised or alternative amplicons designed. For the slope to be an indicator of real amplification (rather than signal drift), there has to be an inflection point.
This is the point on the growth curve when the log-linear phase begins. It also represents the greatest rate of change along the growth curve. (Signal drift is characterised by gradual increase or decrease in fluorescence without amplification of the product.) The important parameter for quantitation is the CT. The higher the initial amount of genomic DNA, the sooner accumulated product is detected in the PCR process, and the lower the CT
value. The threshold should be placed above any baseline activity and within the exponential increase phase (which looks linear in the log transformation). Some software allows determination of the cycle threshold (CT) by a mathematical analysis of the growth curve. This provides better run-to-run reproducibility. A CT value of 40 means no amplification and this value cannot be included in the calculations. Besides being used for quantitation, the CT value can be used for qualitative analysis as a pass/fail measure.
Multiplex TaqMan assays can be performed using multiple dyes with distinct emission wavelengths. Available dyes for this purpose are FAM, TET, VIC and JOE (the most expensive). TAMRA is reserved as the quencher on the probe and ROX as the passive reference. For best results, the combination of FAM (target) and VIC
(endogenous control) is recommended (they have the largest difference in emission maximum) whereas JOE and VIC should not be combined. It is important that if the dye layer has not been chosen correctly, the machine will still read the other dye's spectrum. For example, both VIC and FAM emit fluorescence in a similar range to each other and when doing a single dye, the wells should be labelled correctly. In the case of multiplexing, the spectral compensation for the post run analysis should be turned on (on ABI 7700:
Instrument/Diagnostics/Advanced Options/Miscellaneous). Activating spectral compensation improves dye spectral resolution.
Nested PCR
The disclosed methods can further utilize nested PCR. Nested PCR increases the specificity of DNA amplification, by reducing background due to non-specific amplification of DNA. Two sets of primers are being used in two successive PCRs. In the first reaction, one pair of primers is used to generate DNA products, which besides the intended target, may still consist of non-specifically amplified DNA fragments. The product(s) are then used in a second PCR with a set of primers whose binding sites are completely or partially different from and located 3' of each of the primers used in the first reaction. Nested PCR is often more successful in specifically amplifying long DNA fragments than conventional PCR, but it requires more detailed knowledge of the target sequences.
Primers and Probes The disclosed methods and assays can use primers and probes. As used herein, "primers" are a subset of probes which are capable of supporting some type of enzymatic manipulation and which can hybridize with a target nucleic acid such that the enzymatic manipulation can occur. A primer can be made from any combination of nucleotides or nucleotide derivatives or analogs available in the art which do not interfere with the enzymatic manipulation.
As used herein, "probes" are molecules capable of interacting with a target nucleic acid, typically in a sequence specific manner, for example through hybridization. The hybridization of nucleic acids is well understood in the art and discussed herein. Typically a probe can be made from any combination of nucleotides or nucleotide derivatives or analogs available in the art.
Disclosed are assays and methods which include the use of primers and probes, as well as, the disclosed primer panels all of which are capable of interacting with the disclosed nucleic acids such as ALK (SEQ ID NO: 1), BRAF, EGFR, KIT, or KRAS
or their complement. For example, any of the primers or primer sets from Table 7-14 can be used in the disclosed primer panels or any of the methods and assays disclosed herein.
In certain embodiments the primers are used to support nucleic acid extension reactions, nucleic acid replication reactions, and/or nucleic acid amplification reactions. Typically the primers will be capable of being extended in a sequence specific manner. Extension of a primer in a sequence specific manner includes any methods wherein the sequence and/or composition of the nucleic acid molecule to which the primer is hybridized or otherwise associated directs or influences the composition or sequence of the product produced by the extension of the primer. Extension of the primer in a sequence specific manner therefore includes, but is not limited to, PCR, DNA sequencing, DNA extension, DNA polymerization, RNA
transcription, or reverse transcription. Techniques and conditions that amplify the primer in a sequence specific manner are disclosed. In certain embodiments the primers are used for the DNA amplification reactions, such as PCR or direct sequencing. It is understood that in certain embodiments the primers can also be extended using non-enzymatic techniques, where for example, the nucleotides or oligonucleotides used to extend the primer are modified such that they will chemically react to extend the primer in a sequence specific manner. Typically the disclosed primers hybridize with the disclosed nucleic acids or region of the nucleic acids or they hybridize with the complement of the nucleic acids or complement of a region of the nucleic acids. As an example of the use of primers, one or more primers can be used to create extension products from and templated by a first nucleic acid.
The size of the primers or probes for interaction with the nucleic acids can be any size that supports the desired enzymatic manipulation of the primer, such as DNA
amplification or the simple hybridization of the probe or primer. A typical primer or probe would be at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.
In other embodiments a primer or probe can be less than or equal to 6, 7, 8, 9, 10,

11, 12 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.
The primers for the nucleic acid of interest typically will be used to produce extension products and/or other replicated or amplified products that contain a region of the nucleic acid of interest. The size of the product can be such that the size can be accurately determined to within 3, or 2 or 1 nucleotides.
In certain embodiments the product can be, for example, at least 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.
In other embodiments the product can be, for example, less than or equal to 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, or 4000 nucleotides long.
It is understood and herein contemplated that there are situations where it may be advantageous to utilize more than one primer pair to detect the presence of mutations conferring inhibitor resistance in EGFR, BRAF, KIT, KRAS, or ALK. Such RT-PCR, real-time PCT or other PCR reactions can be conducted separately, or in a single reaction.
When multiple primer pairs are placed into a single reaction, this is referred to as "multiplex PCR." It is understood and herein contemplated that any combination of two or more or three or more the forward and/or reverse primers disclosed herein can be used in the multiplex reaction.
Fluorescent Change Probes and Primers Fluorescent change probes and fluorescent change primers refer to all probes and primers that involve a change in fluorescence intensity or wavelength based on a change in the form or conformation of the probe or primer and nucleic acid to be detected, assayed or replicated. Examples of fluorescent change probes and primers include molecular beacons, Amplifluors, FRET probes, cleavable FRET probes, TaqMan probes, scorpion primers, fluorescent triplex oligos including but not limited to triplex molecular beacons or triplex FRET probes, fluorescent water-soluble conjugated polymers, PNA probes and QPNA
probes.
Fluorescent change probes and primers can be classified according to their structure and/or function. Fluorescent change probes include hairpin quenched probes, cleavage quenched probes, cleavage activated probes, and fluorescent activated probes.
Fluorescent change primers include stem quenched primers and hairpin quenched primers.
Hairpin quenched probes are probes that when not bound to a target sequence form a hairpin structure (and, typically, a loop) that brings a fluorescent label and a quenching moiety into proximity such that fluorescence from the label is quenched. When the probe binds to a target sequence, the stem is disrupted, the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases. Examples of hairpin quenched probes are molecular beacons, fluorescent triplex oligos, triplex molecular beacons, triplex FRET probes, and QPNA probes.
Cleavage activated probes are probes where fluorescence is increased by cleavage of the probe. Cleavage activated probes can include a fluorescent label and a quenching moiety in proximity such that fluorescence from the label is quenched. When the probe is clipped or digested (typically by the 5'-3' exonuclease activity of a polymerase during amplification), the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases. TaqMan probes are an example of cleavage activated probes.
Cleavage quenched probes are probes where fluorescence is decreased or altered by cleavage of the probe. Cleavage quenched probes can include an acceptor fluorescent label and a donor moiety such that, when the acceptor and donor are in proximity, fluorescence resonance energy transfer from the donor to the acceptor causes the acceptor to fluoresce.
The probes are thus fluorescent, for example, when hybridized to a target sequence. When the probe is clipped or digested (typically by the 5'-3' exonuclease activity of a polymerase during amplification), the donor moiety is no longer in proximity to the acceptor fluorescent label and fluorescence from the acceptor decreases. If the donor moiety is itself a fluorescent label, it can release energy as fluorescence (typically at a different wavelength than the fluorescence of the acceptor) when not in proximity to an acceptor.
The overall effect would then be a reduction of acceptor fluorescence and an increase in donor fluorescence. Donor fluorescence in the case of cleavage quenched probes is equivalent to fluorescence generated by cleavage activated probes with the acceptor being the quenching moiety and the donor being the fluorescent label. Cleavable FRET (fluorescence resonance energy transfer) probes are an example of cleavage quenched probes.
Fluorescent activated probes are probes or pairs of probes where fluorescence is increased or altered by hybridization of the probe to a target sequence.
Fluorescent activated probes can include an acceptor fluorescent label and a donor moiety such that, when the acceptor and donor are in proximity (when the probes are hybridized to a target sequence), fluorescence resonance energy transfer from the donor to the acceptor causes the acceptor to fluoresce. Fluorescent activated probes are typically pairs of probes designed to hybridize to adjacent sequences such that the acceptor and donor are brought into proximity.
Fluorescent activated probes can also be single probes containing both a donor and acceptor where, when the probe is not hybridized to a target sequence, the donor and acceptor are not in proximity but where the donor and acceptor are brought into proximity when the probe hybridized to a target sequence. This can be accomplished, for example, by placing the donor and acceptor on opposite ends of the probe and placing target complement sequences at each end of the probe where the target complement sequences are complementary to adjacent sequences in a target sequence. If the donor moiety of a fluorescent activated probe is itself a fluorescent label, it can release energy as fluorescence (typically at a different wavelength than the fluorescence of the acceptor) when not in proximity to an acceptor (that is, when the probes are not hybridized to the target sequence).
When the probes hybridize to a target sequence, the overall effect would then be a reduction of donor fluorescence and an increase in acceptor fluorescence. FRET probes are an example of fluorescent activated probes.
Stem quenched primers are primers that when not hybridized to a complementary sequence form a stem structure (either an intramolecular stem structure or an intermolecular stem structure) that brings a fluorescent label and a quenching moiety into proximity such that fluorescence from the label is quenched. When the primer binds to a complementary sequence, the stem is disrupted; the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases. In the disclosed method, stem quenched primers are used as primers for nucleic acid synthesis and thus become incorporated into the synthesized or amplified nucleic acid. Examples of stem quenched primers are peptide nucleic acid quenched primers and hairpin quenched primers.
Peptide nucleic acid quenched primers are primers associated with a peptide nucleic acid quencher or a peptide nucleic acid fluor to form a stem structure. The primer contains a fluorescent label or a quenching moiety and is associated with either a peptide nucleic acid quencher or a peptide nucleic acid fluor, respectively. This puts the fluorescent label in proximity to the quenching moiety. When the primer is replicated, the peptide nucleic acid is displaced, thus allowing the fluorescent label to produce a fluorescent signal.
Hairpin quenched primers are primers that when not hybridized to a complementary sequence form a hairpin structure (and, typically, a loop) that brings a fluorescent label and a quenching moiety into proximity such that fluorescence from the label is quenched. When the primer binds to a complementary sequence, the stem is disrupted; the quenching moiety is no longer in proximity to the fluorescent label and fluorescence increases.
Hairpin quenched primers are typically used as primers for nucleic acid synthesis and thus become incorporated into the synthesized or amplified nucleic acid. Examples of hairpin quenched primers are Amplifluor primers and scorpion primers.
Cleavage activated primers are similar to cleavage activated probes except that they are primers that are incorporated into replicated strands and are then subsequently cleaved.

Labels To aid in detection and quantitation of nucleic acids produced using the disclosed methods, labels can be directly incorporated into nucleotides and nucleic acids or can be coupled to detection molecules such as probes and primers. As used herein, a label is any molecule that can be associated with a nucleotide or nucleic acid, directly or indirectly, and which results in a measurable, detectable signal, either directly or indirectly. Many such labels for incorporation into nucleotides and nucleic acids or coupling to nucleic acid probes are known to those of skill in the art. Examples of labels suitable for use in the disclosed method are radioactive isotopes, fluorescent molecules, phosphorescent molecules, enzymes, antibodies, and ligands. Fluorescent labels, especially in the context of fluorescent change probes and primers, are useful for real-time detection of amplification.
Examples of suitable fluorescent labels include fluorescein isothiocyanate (FITC), 5,6-carboxymethyl fluorescein, Texas red, nitrobenz-2-oxa-1,3-diazol-4-y1 (NBD), coumarin, dansyl chloride, rhodamine, amino-methyl coumarin (AMCA), Eosin, Erythrosin, BODIPY , CASCADE BLUE , OREGON GREEN , pyrene, lissamine, xanthenes, acridines, oxazines, phycoerythrin, macrocyclic chelates of lanthanide ions such as quantum dyeTM, fluorescent energy transfer dyes, such as thiazole orange-ethidium heterodimer, and the cyanine dyes Cy3, Cy3.5, Cy5, Cy5.5 and Cy7. Examples of other specific fluorescent labels include 3-Hydroxypyrene 5,8,10-Tri Sulfonic acid, 5-Hydroxy Tryptamine (5-HT), Acid Fuchsin, Alizarin Complexon, Alizarin Red, Allophycocyanin, Aminocoumarin, Anthroyl Stearate, Astrazon Brilliant Red 4G, Astrazon Orange R, Astrazon Red 6B, Astrazon Yellow 7 GLL, Atabrine, Auramine, Aurophosphine, Aurophosphine G, BAO

(Bisaminophenyloxadiazole), BCECF, BerberineSulphate, Bisbenzamide, Blancophor FFG
Solution, Blancophor SV, Bodipy Fl, Brilliant Sulphoflavin FF, Calcien Blue, Calcium Green, Calcofluor RW Solution, Calcofluor White, Calcophor White ABT Solution, Calcophor White Standard Solution, Carbostyryl, Cascade Yellow, Catecholamine, Chinacrine, Coriphosphine 0, Coumarin-Phalloidin, CY3.1 8, CY5.1 8, CY7, Dans (1-Dimethyl Amino Naphaline 5 Sulphonic Acid), Dansa (DiaminoNaphtylSulphonic Acid), Dansyl NH-CH3, Diamino Phenyl Oxydiazole (DAO), Dimethylamino-5-Sulphonic acid, DipyrrometheneboronDifluoride, Diphenyl Brilliant Flavine 7GFF, Dopamine, Erythrosin ITC, Euchrysin, FIF (Formaldehyde Induced Fluorescence), Flazo Orange, Fluo 3, Fluorescamine, Fura-2, Genacryl Brilliant Red B, Genacryl Brilliant Yellow 10GF, Genacryl Pink 3G, Genacryl Yellow 5GF, Gloxalic Acid, Granular Blue, Haematoporphyrin, Indo-1, IntrawhiteCf Liquid, Leucophor PAF, Leucophor SF, Leucophor WS, LissamineRhodamine B200 (RD200), Lucifer Yellow CH, Lucifer Yellow VS, Magdala Red, Marina Blue, Maxilon Brilliant Flavin 10 GFF, Maxilon Brilliant Flavin 8 GFF, MPS (Methyl Green PyronineStilbene), Mithramycin, NBD Amine, Nitrobenzoxadidole, Noradrenaline, Nuclear Fast Red, Nuclear Yellow, Nylosan Brilliant Flavin E8G, Oxadiazole, Pacific Blue, Pararosaniline (Feulgen), Phorwite AR
Solution, Phorwite BKL, Phorwite Rev, Phorwite RPA, Phosphine 3R, Phthalocyanine, Phycoerythrin R, Phycoerythrin B, PolyazaindacenePontochrome Blue Black, Porphyrin, Primuline, Procion Yellow, Pyronine, Pyronine B, Pyrozal Brilliant Flavin 7GF, Quinacrine Mustard, Rhodamine 123, Rhodamine 5 GLD, Rhodamine 6G, Rhodamine B, Rhodamine B
200, Rhodamine B Extra, Rhodamine BB, Rhodamine BG, Rhodamine WT, Serotonin, Sevron Brilliant Red 2B, Sevron Brilliant Red 4G, Sevron Brilliant Red B, Sevron Orange, Sevron Yellow L, SITS (Primuline), SITS (StilbeneIsothiosulphonic acid), Stilbene, Snarf 1, sulphoRhodamine B Can C, SulphoRhodamine G Extra, Tetracycline, Thiazine Red R, Thioflavin S, Thioflavin TCN, Thioflavin 5, Thiolyte, Thiozol Orange, Tinopol CBS, True Blue, Ultralite, Uranine B, Uvitex SFC, Xylene Orange, and XRITC.
The absorption and emission maxima, respectively, for some of these fluors are:
FITC (490 nm; 520 nm), Cy3 (554 nm; 568 nm), Cy3.5 (581 nm; 588 nm), Cy5 (652 nm:
672 nm), Cy5.5 (682 nm; 703 nm) and Cy7 (755 nm; 778 nm), thus allowing their simultaneous detection. Other examples of fluorescein dyes include 6-carboxyfluorescein (6-FAM), 2',4',1,4,-tetrachlorofluorescein (TET), 2',4',5',7',1,4-hexachlorofluorescein (HEX), 2',7'-dimethoxy-4', 5'-dichloro-6-carboxyrhodamine (JOE), 2'-chloro-5'-fluoro-7',8'-fused phenyl-1,4-dichloro-6-carboxyfluorescein (NED), and 2'-chloro-7'-pheny1-1,4-dichloro-6-carboxyfluorescein (VIC). Fluorescent labels can be obtained from a variety of commercial sources, including Amersham Pharmacia Biotech, Piscataway, NJ;
Molecular Probes, Eugene, OR; and Research Organics, Cleveland, Ohio.
Additional labels of interest include those that provide for signal only when the probe with which they are associated is specifically bound to a target molecule, where such labels include: "molecular beacons" as described in Tyagi& Kramer, Nature Biotechnology (1996) 14:303 and EP 0 070 685 Bl. Other labels of interest include those described in U.S. Pat. No. 5,563,037 which is incorporated herein by reference.
Labeled nucleotides are a form of label that can be directly incorporated into the amplification products during synthesis. Examples of labels that can be incorporated into amplified nucleic acids include nucleotide analogs such as BrdUrd, aminoallyldeoxyuridine, 5-methylcytosine, bromouridine, and nucleotides modified with biotin or with suitable haptens such as digoxygenin. Suitable fluorescence-labeled nucleotides are Fluorescein-isothiocyanate-dUTP, Cyanine-3-dUTP and Cyanine-5-dUTP. One example of a nucleotide analog label for DNA is BrdUrd (bromodeoxyuridine, BrdUrd, BrdU, BUdR, Sigma-Aldrich Co). Other examples of nucleotide analogs for incorporation of label into DNA are AA-dUTP (aminoallyl-deoxyuridine triphosphate, Sigma-Aldrich Co.), and 5-methyl-dCTP
(Roche Molecular Biochemicals). One example of a nucleotide analog for incorporation of label into RNA is biotin-16-UTP (biotin-16-uridine-5'-triphosphate, Roche Molecular Biochemicals). Fluorescein, Cy3, and Cy5 can be linked to dUTP for direct labeling.
Cy3.5 and Cy7 are available as avidin or anti-digoxygenin conjugates for secondary detection of biotin- or digoxygenin-labeled probes.
Labels that are incorporated into amplified nucleic acid, such as biotin, can be subsequently detected using sensitive methods well-known in the art. For example, biotin can be detected using streptavidin-alkaline phosphatase conjugate (Tropix, Inc.), which is bound to the biotin and subsequently detected by chemiluminescence of suitable substrates (for example, chemiluminescent substrate CSPD: disodium, 3-(4-methoxyspiro-[1,2,-dioxetane-3-2'-(5'-chloro)tricyclo [3.3.1.1ndecane]-4-y1) phenyl phosphate;
Tropix, Inc.).

Labels can also be enzymes, such as alkaline phosphatase, soybean peroxidase, horseradish peroxidase and polymerases, that can be detected, for example, with chemical signal amplification or by using a substrate to the enzyme which produces light (for example, a chemiluminescent 1,2-dioxetane substrate) or fluorescent signal.
Molecules that combine two or more of these labels are also considered labels.
Any of the known labels can be used with the disclosed probes, tags, and method to label and detect nucleic acid amplified using the disclosed method. Methods for detecting and measuring signals generated by labels are also known to those of skill in the art. For example, radioactive isotopes can be detected by scintillation counting or direct visualization; fluorescent molecules can be detected with fluorescent spectrophotometers;
phosphorescent molecules can be detected with a spectrophotometer or directly visualized with a camera; enzymes can be detected by detection or visualization of the product of a reaction catalyzed by the enzyme; antibodies can be detected by detecting a secondary label coupled to the antibody. As used herein, detection molecules are molecules which interact with amplified nucleic acid and to which one or more labels are coupled.
The disclosed methods, assays, and primer panels can be used to diagnose any disease where uncontrolled cellular proliferation occurs herein referred to as "cancer". A
non-limiting list of different types of ALK related cancers is as follows:
lymphomas (Hodgkins and non-Hodgkins), leukemias, carcinomas, carcinomas of solid tissues, squamous cell carcinomas, adenocarcinomas, sarcomas, gliomas, high grade gliomas, blastomas, neuroblastomas, plasmacytomas, histiocytomas, melanomas, adenomas, hypoxic tumours, myelomas, AIDS-related lymphomas or sarcomas, metastatic cancers, or cancers in general. In particular, the disclosed methods, assays, and kits relate to the diagnosis, detection, or prognosis of inflammatory breast cancer A representative but non-limiting list of cancers that the disclosed methods can be used to diagnose is the following: lymphoma, B cell lymphoma (including diffuse large B-cell lymphoma), B-cell plasmablastic/immunoblastic lymphomas, T cell lymphoma (including T- or null-cell lymphomas), mycosis fungoides, Hodgkin's Disease, myeloid leukemia, bladder cancer, brain cancer, nervous system cancer, head and neck cancer, squamous cell carcinoma of head and neck, kidney cancer, lung cancers such as small cell lung cancer and non-small cell lung cancer, neuroblastoma/glioblastomaõ anaplastic large-cell lymphoma (ALCL), inflammatory myofibroblastic tumors, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, liver cancer, melanoma, malignant histiocytosis, squamous cell carcinomas of the mouth, throat, larynx, and lung, colon cancer, cervical cancer, cervical carcinoma, breast cancer (including inflammatory breast cancer), and epithelial cancer, renal cancer, genitourinary cancer, pulmonary cancer, esophageal squamous cell carcinoma, head and neck carcinoma, large bowel cancer, hematopoietic cancers; testicular cancer;
colon and rectal cancers, prostatic cancer, or pancreatic cancer.
Nucleic Acids The disclosed method and compositions make use of various nucleic acids.
Generally, any nucleic acid can be used in the disclosed method. For example, the disclosed nucleic acids of interest and the disclosed reference nucleic acids can be chosen based on the desired analysis and information that is to be obtained or assessed. The disclosed methods also produce new and altered nucleic acids. The nature and structure of such nucleic acids will be established by the manner in which they are produced and manipulated in the methods. Thus, for example, extension products and hybridizing nucleic acids are produced in the disclosed methods. As used herein, hybridizing nucleic acids are hybrids of extension products and the second nucleic acid.
It is understood and contemplated herein that a nucleic acid of interest can be any nucleic acid to which the determination of the presence or absence of nucleotide variation is desired. Thus, for example, the nucleic acid of interest can comprise a sequence that corresponds to the wild-type sequence of the reference nucleic acid. It is further disclosed herein that the disclosed methods can be performed where the first nucleic acid is a reference nucleic acid and the second nucleic acid is a nucleic acid of interest or where the first nucleic acid is the nucleic acid of interest and the second nucleic acid is the reference nucleic acid.

It is understood and herein contemplated that a reference nucleic acid can be any nucleic acid against which a nucleic acid of interest is to be compared.
Typically, the reference nucleic acid has a known sequence (and/or is known to have a sequence of interest as a reference). Although not required, it is useful if the reference sequence has a known or suspected close relationship to the nucleic acid of interest. For example, if a single nucleotide variation is desired to be detected, the reference sequence can be usefully chosen to be a sequence that is a homolog or close match to the nucleic acid of interest, such as a nucleic acid derived from the same gene or genetic element from the same or a related organism or individual. Thus, for example, it is contemplated herein that the reference nucleic acid can comprise a wild-type sequence or alternatively can comprise a known mutation including, for example, a mutation the presence or absence of which is associated with a disease or resistance to therapeutic treatment. Thus, for example, it is contemplated that the disclosed methods can be used to detect or diagnose the presence of known mutations in a nucleic acid of interest by comparing the nucleic acid of interest to a reference nucleic acid that comprises a wild-type sequence (i.e., is known not to possess the mutation) and examining for the presence or absence of variation in the nucleic acid of interest, where the absence of variation would indicate the absence of a mutation.
Alternatively, the reference nucleic acid can possess a known mutation. Thus, for example, it is contemplated that the disclosed methods can be used to detect susceptibility for a disease or condition by comparing the nucleic acid of interest to a reference nucleic acid comprising a known mutation that indicates susceptibility for a disease and examining for the presence or absence of the mutation, wherein the presence of the mutation indicates a disease.
Herein, the term "nucleotide variation" refers to any change or difference in the nucleotide sequence of a nucleic acid of interest relative to the nucleotide sequence of a reference nucleic acid. Thus, a nucleotide variation is said to occur when the sequences between the reference nucleic acid and the nucleic acid of interest (or its complement, as appropriate in context) differ. Thus, for example, a substitution of an adenine (A) to a guanine (G) at a particular position in a nucleic acid would be a nucleotide variation provided the reference nucleic acid comprised an A at the corresponding position. It is understood and herein contemplated that the determination of a variation is based upon the reference nucleic acid and does not indicate whether or not a sequence is wild-type. Thus, for example, when a nucleic acid with a known mutation is used as the reference nucleic acid, a nucleic acid not possessing the mutation (including a wild-type nucleic acid) would be considered to possess a nucleotide variation (relative to the reference nucleic acid).
Nucleotides The disclosed methods and compositions make use of various nucleotides.
Throughout this application and the methods disclosed herein reference is made to the type of base for a nucleotide. It is understood and contemplated herein that where reference is made to a type of base, this refers a base that in a nucleotide in a nucleic acid strand is capable of hybridizing (binding) to a defined set of one or more of the canonical bases.
Thus, for example, where reference is made to extension products extended in the presence of three types of nuclease resistant nucleotides and not in the presence of nucleotides that comprise the same type of base as the modified nucleotides, this means that if, for example, the base of the modified nucleotide was an adenine (A), the nuclease-resistant nucleotides can be, for example, guanine (G), thymine (T), and cytosine (C). Each of these bases (which represent the four canonical bases) is capable of hybridizing to a different one of the four canonical bases and thus each qualify as a different type of base as defined herein. As another example, inosine base pairs primarily with adenine and cytosine (in DNA) and thus can be considered a different type of base from adenine and from cytosine-which base pair with thymine and guanine, respectively-but not a different type of base from guanine or thymine-which base pair with cytosine and adenine, respectively-because the base pairing of guanine and thymine overlaps (that is, is not different from) the hybridization pattern of inosine.
Any type of modified or alternative base can be used in the disclosed methods and compositions, generally limited only by the capabilities of the enzymes used to use such bases. Many modified and alternative nucleotides and bases are known, some of which are described below and elsewhere herein. The type of base that such modified and alternative bases represent can be determined by the pattern of base pairing for that base as described herein. Thus for example, if the modified nucleotide was adenine, any analog, derivative, modified, or variant base that based pairs primarily with thymine would be considered the same type of base as adenine. In other words, so long as the analog, derivative, modified, or variant has the same pattern of base pairing as another base, it can be considered the same type of base. Modifications can made to the sugar or phosphate groups of a nucleotide.
Generally such modifications will not change the base pairing pattern of the base.
However, the base pairing pattern of a nucleotide in a nucleic acid strand determines the type of base of the base in the nucleotide.
Modified nucleotides to be incorporated into extension products and to be selectively removed by the disclosed agents in the disclosed methods can be any modified nucleotide that functions as needed in the disclosed method as is described elsewhere herein. Modified nucleotides can also be produced in existing nucleic acid strands, such as extension products, by, for example, chemical modification, enzymatic modification, or a combination.
Many types of nuclease-resistant nucleotides are known and can be used in the disclosed methods. For example, nucleotides have modified phosphate groups and/or modified sugar groups can be resistant to one or more nucleases. Nuclease-resistance is defined herein as resistance to removal from a nucleic acid by any one or more nucleases.
Generally, nuclease resistance of a particular nucleotide can be defined in terms of a relevant nuclease. Thus, for example, if a particular nuclease is used in the disclosed method, the nuclease-resistant nucleotides need only be resistant to that particular nuclease.
Examples of useful nuclease-resistant nucleotides include thio-modified nucleotides and borano-modified nucleotides.
There are a variety of molecules disclosed herein that are nucleic acid based.
Non-limiting examples of these and other molecules are discussed herein. It is understood that for example, a nucleotide is a molecule that contains a base moiety, a sugar moiety and a phosphate moiety. Nucleotides can be linked together through their phosphate moieties and sugar moieties creating an intemucleoside linkage. The base moiety of a nucleotide can be adenine-9-y1 (adenine, A), cytosine-1-y' (cytosine, C), guanine-9-y1 (guanine, G), uracil-1-yl (uracil, U), and thymin-l-yl (thymine, T). The sugar moiety of a nucleotide is a ribose or a deoxyribose. The phosphate moiety of a nucleotide is pentavalent phosphate.
A non-limiting example of a nucleotide would be 3'-AMP (3'-adenosine monophosphate) or 5'-GMP (5'-guanosine monophosphate).
A nucleotide analog is a nucleotide which contains some type of modification to either the base, sugar, or phosphate moieties. Modifications to the base moiety would include natural and synthetic modifications of A, C, G, and T/U as well as different purine or pyrimidine bases, such as uracil-5-y1 (y), hypoxanthin-9-y1 (inosine, I), and 2-aminoadenin-9-yl. A modified base includes but is not limited to 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.
Additional base modifications can be found for example in U.S. Pat. No. 3,687,808, which is incorporated herein in its entirety for its teachings of base modifications.
Certain nucleotide analogs, such as 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine can increase the stability of duplex formation. Often time base modifications can be combined with for example a sugar modification, such as 2'-0-methoxyethyl, to achieve unique properties such as increased duplex stability.

Nucleotide analogs can also include modifications of the sugar moiety.
Modifications to the sugar moiety would include natural modifications of the ribose and deoxy ribose as well as synthetic modifications. Sugar modifications include but are not limited to the following modifications at the 2' position: OH; F; 0-, S-, or N-alkyl; 0-, S-, or N-alkenyl; 0-, S-or N-alkynyl; or 0-alkyl-0-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted Cl to C10, alkyl or C2 to C10 alkenyl and alkynyl. 2' sugar modifications also include but are not limited to -0[(CH2)n O]m CH3, -0(CH2)n OCH3, -0(CH2)n NH2, -0(CH2)n CH3, -0(CH2)n -ONH2, and -0(CH2)nON[(CH2)n CH3)]2, where n and m are from 1 to about 10.
Other modifications at the 2' position include but are not limited to: Cl to C10 lower alkyl, substituted lower alkyl, alkaryl, aralkyl, 0-alkaryl or 0-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2 CH3, 0NO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA
cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an oligonucleotide, or a group for improving the pharmacodynamic properties of an oligonucleotide, and other substituents having similar properties.
Similar modifications may also be made at other positions on the sugar, particularly the 3' position of the sugar on the 3' terminal nucleotide or in 2'-5' linked oligonucleotides and the 5' position of 5' terminal nucleotide. Modified sugars would also include those that contain modifications at the bridging ring oxygen, such as CH2 and S. Nucleotide sugar analogs may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar.
Nucleotide analogs can also be modified at the phosphate moiety. Modified phosphate moieties include but are not limited to those that can be modified so that the linkage between two nucleotides contains a phosphorothioate, chiral phosphorothioate, phosphorodithioate, phosphotriester, aminoalkylphosphotriester, methyl and other alkyl phosphonates including 3'-alkylene phosphonate and chiral phosphonates, phosphinates, phosphoramidates including 3'-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates. It is understood that these phosphate or modified phosphate linkage between two nucleotides can be through a 3'-5' linkage or a 2'-5' linkage, and the linkage can contain inverted polarity such as 3'-5' to 5'-3' or 2'-5' to 5'-2'.
Various salts, mixed salts and free acid forms are also included.
It is understood that nucleotide analogs need only contain a single modification, but may also contain multiple modifications within one of the moieties or between different moieties.
Nucleotide substitutes are molecules having similar functional properties to nucleotides, but which do not contain a phosphate moiety, such as peptide nucleic acid (PNA). Nucleotide substitutes are molecules that will recognize nucleic acids in a Watson-Crick or Hoogsteen manner, but which are linked together through a moiety other than a phosphate moiety. Nucleotide substitutes are able to conform to a double helix type structure when interacting with the appropriate target nucleic acid.
Nucleotide substitutes are nucleotides or nucleotide analogs that have had the phosphate moiety and/or sugar moieties replaced. Nucleotide substitutes do not contain a standard phosphorus atom. Substitutes for the phosphate can be for example, short chain alkyl or cycloalkylintemucleoside linkages, mixed heteroatom and alkyl or cycloalkylinternucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones;
methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones;
amide backbones; and others having mixed N, 0, S and CH2 component parts.
It is also understood in a nucleotide substitute that both the sugar and the phosphate moieties of the nucleotide can be replaced, by for example an amide type linkage (aminoethylglycine) (PNA).

It is also possible to link other types of molecules (conjugates) to nucleotides or nucleotide analogs. Conjugates can be chemically linked to the nucleotide or nucleotide analogs. Such conjugates include but are not limited to lipid moieties such as a cholesterol moiety, cholic acid, a thioether, e.g., hexyl-S-tritylthiol, a thiocholesterol, an aliphatic chain, e.g., dodecandiol or undecyl residues, a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate, a polyamine or a polyethylene glycol chain, or adamantane acetic acid, a palmityl moiety, or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety.
A Watson-Crick interaction is at least one interaction with the Watson-Crick face of a nucleotide, nucleotide analog, or nucleotide substitute. The Watson-Crick face of a nucleotide, nucleotide analog, or nucleotide substitute includes the C2, Ni, and C6 positions of a purine based nucleotide, nucleotide analog, or nucleotide substitute and the C2, N3, C4 positions of a pyrimidine based nucleotide, nucleotide analog, or nucleotide substitute.
A Hoogsteen interaction is the interaction that takes place on the Hoogsteen face of a nucleotide or nucleotide analog, which is exposed in the major groove of duplex DNA.
The Hoogsteen face includes the N7 position and reactive groups (NH2 or 0) at the C6 position of purine nucleotides.
Hybridization/Selective Hybridization The term hybridization typically means a sequence driven interaction between at least two nucleic acid molecules, such as a primer or a probe and a gene.
Sequence driven interaction means an interaction that occurs between two nucleotides or nucleotide analogs or nucleotide derivatives in a nucleotide specific manner. For example, G
interacting with C or A interacting with T are sequence driven interactions. Typically sequence driven interactions occur on the Watson-Crick face or Hoogsteen face of the nucleotide. The hybridization of two nucleic acids is affected by a number of conditions and parameters known to those of skill in the art. For example, the salt concentrations, pH, and temperature of the reaction all affect whether two nucleic acid molecules will hybridize.

Parameters for selective hybridization between two nucleic acid molecules are well known to those of skill in the art. For example, in some embodiments selective hybridization conditions can be defined as stringent hybridization conditions.
For example, stringency of hybridization is controlled by both temperature and salt concentration of either or both of the hybridization and washing steps. For example, the conditions of hybridization to achieve selective hybridization may involve hybridization in high ionic strength solution (6X SSC or 6X SSPE) at a temperature that is about 12-25 C
below the Tm (the melting temperature at which half of the molecules dissociate from their hybridization partners) followed by washing at a combination of temperature and salt concentration chosen so that the washing temperature is about 5 C to 20 C
below the Tm.
The temperature and salt conditions are readily determined empirically in preliminary experiments in which samples of reference DNA immobilized on filters are hybridized to a labeled nucleic acid of interest and then washed under conditions of different stringencies.
Hybridization temperatures are typically higher for DNA-RNA and RNA-RNA
hybridizations. The conditions can be used as described above to achieve stringency, or as is known in the art. A preferable stringent hybridization condition for a DNA:DNA
hybridization can be at about 68 C (in aqueous solution) in 6X SSC or 6X SSPE
followed by washing at 68 C. Stringency of hybridization and washing, if desired, can be reduced accordingly as the degree of complementarity desired is decreased, and further, depending upon the G-C or A-T richness of any area wherein variability is searched for.
Likewise, stringency of hybridization and washing, if desired, can be increased accordingly as homology desired is increased, and further, depending upon the G-C or A-T
richness of any area wherein high homology is desired, all as known in the art.
Another way to define selective hybridization is by looking at the amount (percentage) of one of the nucleic acids bound to the other nucleic acid. For example, in some embodiments selective hybridization conditions would be when at least about, 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 percent of the limiting nucleic acid is bound to the non-limiting nucleic acid. Typically, the non-limiting primer is in for example, 10 or 100 or 1000 fold excess. This type of assay can be performed at under conditions where both the limiting and non-limiting primer are for example, 10 fold or 100 fold or 1000 fold below their kd, or where only one of the nucleic acid molecules is 10 fold or 100 fold or 1000 fold or where one or both nucleic acid molecules are above their kd.
Another way to define selective hybridization is by looking at the percentage of primer that gets enzymatically manipulated under conditions where hybridization is required to promote the desired enzymatic manipulation. For example, in some embodiments selective hybridization conditions would be when at least about, 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 percent of the primer is enzymatically manipulated under conditions which promote the enzymatic manipulation, for example if the enzymatic manipulation is DNA extension, then selective hybridization conditions would be when at least about 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 percent of the primer molecules are extended.
Conditions also include those suggested by the manufacturer or indicated in the art as being appropriate for the enzyme performing the manipulation.
Just as with homology, it is understood that there are a variety of methods herein disclosed for determining the level of hybridization between two nucleic acid molecules. It is understood that these methods and conditions may provide different percentages of hybridization between two nucleic acid molecules, but unless otherwise indicated meeting the parameters of any of the methods would be sufficient. For example if 80%
hybridization was required and as long as hybridization occurs within the required parameters in any one of these methods it is considered disclosed herein.
It is understood that those of skill in the art understand that if a composition or method meets any one of these criteria for determining hybridization either collectively or singly it is a composition or method that is disclosed herein.
Kits Disclosed herein are kits that are drawn to reagents that can be used in practicing the methods disclosed herein. In particular, he kits can include any reagent or combination of reagents discussed herein or that would be understood to be required or beneficial in the practice of the disclosed methods. For example, the kits could include one or more primers from Tables 7-14 disclosed herein to perform the extension, replication and amplification reactions discussed in certain embodiments of the methods, as well as the buffers and enzymes required to use the primers as intended. The kit can also include other necessary reagents to perform any of the next generation sequencing techniques disclosed herein. In another aspect, the disclosed kits can include one or more of the probes listed in Table 15 in addition to or instead of one or more primers from Table 7-14.
It is understood and herein contemplated that the disclosed kits can comprise at least one primer set to detect the presence of nucleic acid variation in each of KIT, BRAF, KRAS, ALK, and EGFR. For example, the kits can comprise at least one primer or primer set for sequencing at least one of each of the KIT, BRAF, KRAS, ALK, and EGFR exons of Tables 1. In one aspect, the kits can comprise at least one primer or primer set from each of Tables 7-14. Alternatively, the kit can comprise a primer or primer set that will detect one or more of the specific mutations listed in Tables 2-6. Therefore, in one aspect disclosed herein are kits for performing a NGS sequencing reaction on a tissue sample to detect the presence of a mutation conferring kinase inhibitor resistance comprising at least one or more primer or primer set from each of Table 7-14. In another aspect, disclosed herein are kits for performing a NGS sequencing reaction on a tissue sample to detect the presence of a mutation conferring kinase inhibitor resistance comprising at least one or more primer or primer set capable of specifically hybridizing an amplifying any of the mutant sequences of KIT, BRAF, KRAS, ALK, and EGFR present in Tables 2-6.
Additionally, it is understood that the disclosed kits can include such other reagents and material for performing the disclosed methods such as enzymes (e.g., polymerases), buffers, sterile water, and/or reaction tubes. Additionally the kits can also include modified nucleotides, nuclease-resistant nucleotides, and or labeled nucleotides.
Additionally, the disclosed kits can include instructions for performing the methods disclosed herein and software for enable the calculation of the presence of a kinase inhibitor mutation (i.e., a mutation in KIT, BRAF, KRAS, EGFR, and/or ALK).
The compositions disclosed herein and the compositions necessary to perform the disclosed methods can be made using any method known to those of skill in the art for that particular reagent or compound unless otherwise specifically noted.
Nucleic Acid Synthesis The disclosed nucleic acids, such as, the oligonucleotides to be used as primers can be made using standard chemical synthesis methods or can be produced using enzymatic methods or any other known method. Such methods can range from standard enzymatic digestion followed by nucleotide fragment isolation to purely synthetic methods, for example, by the cyanoethylphosphoramidite method using a Milligen or Beckman System 1Plus DNA synthesizer (for example, Model 8700 automated synthesizer of Milligen-Biosearch, Burlington, MA or ABI Model 380B).
Examples The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and/or methods claimed herein are made and evaluated, and are intended to be purely exemplary and are not intended to limit the disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in C or is at ambient temperature, and pressure is at or near atmospheric.
Example 1: ALK Inhibitor Resistance Using an in vitro assay known to predict clinically relevant kinase inhibitor-resistance mutations resistance selection studies were performed with XALKORIO
and identified a large number of ALK kinase domain point mutations that confer high-level resistance to the Pfizer inhibitor (Figure 1). In response to the issue of resistance, a number of pharma and biotech companies currently have 2n1-generation ALK small-molecule inhibitors in development.
The need for more comprehensive oncogene profiling in patients with ALK
inhibitor resistance was observed in an ALK positive crizotinib resistant cohort of patients that ALK
specific kinase mutations accounted for only a third of crizotinib resistance.
The larger subset of crizotinib resistant cases indicated that second (co-expression in conjunction with ALK) or separate (complete absence of ALK) oncogenic drivers such as EGFR, BRAF, KRAS or cKIT can relieve the sensitivity to crizotinib and drive oncogenesis.
It was also observed in a single case that the complete loss of ALK expression did not correspond to the presence of an identifiable alternate driver indicating the genetic profiling of ALK inhibitor resistance cases should be extended past EGFR, BRAF, KRAS or cKIT expression using more versatile testing platforms. The presence of multiple oncogenes present in a single tumor sample is by no means a new phenomenon as EGFR driven tumors resistant to EGFR
tyrosine kinase inhibitors can be driven by secondary MET gene amplification.
Example 2: A Diagnostic Cancer Panel that Employs NGS
Applicants have designed and developed a next generation sequencing panel to amplify and sequence one or more exons within ALK and other oncogenes implicated in driving tumorigenesis in the presence of crizotinib (i.e. ALK, BRAF, EGFR, KIT
and K-RAS. See Table 1 for an overarching description of the exons targeted for sequencing in the panel and Tables 2-6 for a more detailed list of each mutation detected by the Insight ALK
resistance IDTM panel. Primer sequences used to amplify each gene segment are depicted in Tables 7-14.
Table 1. Exons That Are Covered Gene Exon Example 3: Targeted Next Generation Sequencing Insight ALK resistance IDTM
Polymerase chain reaction is used to create amplicons that span the exonic regions mentioned above. The design described here is agnostic to the NGS platform used to perform the actual sequencing, and thus multiple PCR strategies can match the size of the PCR fragments to the read-length of the sequencing platform being employed.
The PCR
amplification can be done in a single-tube as a multiple reaction where all targets are covered at once. In the case of low coverage or ambiguous results, a single-plex PCR can be performed as a confirmatory step to ensure accurate mutation calling. This is also true in the case of highly-degraded samples where the template DNA has fragmented and large-amplicons cannot be extracted from the DNA that remains. See Tables 7-14 for a full list of the primers that have been designed and the general size of fragments each set produces.
There are a large number of primers in the list to ensure that there is flexibility to run various multiplex PCR reactions where there is very little sequence overlap in the primers, which can lead to dimerization, and allow melting temperatures of all the oligos in a particular reaction to be matched. The amplification parameters of each PCR
reaction consist of 95 C 15-min heat denaturation phase followed by 40 cycles of denaturation at 95 C for 15 sec and 55 C annealing for 30 sec and 72 C extension for 1 min and finally a 72 C final extension step for 5 minutes. At the end of the PCR step a diverse set of fragments that cover the exons of interest can be synthesized. The fragments can then be adapted for sequencing on any commercially available NGS platform. Since there is a very wide range of read-lengths that the different NGS instruments produce, from as low as 35 bases to as high as 1500 and expectations of 100kb read length in the near future, the Insight ALK resistance IDTM is designed to be able to produce fragments as short as 150 bases to as high as 5kb. This ensures for efficient sequencing where the size of each amplicon can be matched to the output of long-read and middle-read technologies (150-1000 bases) or have large enough fragments (5kb) that can be effectively sheared, either sonically or enzymatically, to be compatible with short-read sequencers (<150 bases).
The ALK resistance IDTM takes advantage of the very high-throughput offered by modern sequencers to cover the regions of interest at very high coverage (depth > 5,000X) and thus enable the detection of rare variants only present in the sample at a frequency of 1% or less. The sequence reads that are generated can be compared to a reference sequence examined for the presence of any of the mutations listed in Tables 2-6.
Table 2. ALK Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.V1471fs*45 c.4409 _4422delCCGTGGAAGGGGGA
p.Y1584Y c.4752C>T
p.115971 c.4791T>A
p.L10621 c.3185A>T
p.110871 c.3260C>T
p.D1091N c.3271G>A
p.G1128A c.3383G>C
p.M1166R c.3497T>G
p.A1168P c.3502C>G
p.I1171N c.3512T>A
p.F1174I c.3520T>A
p.F1174L c.3522C>A
p.R1192P c.3575G>C
p.F1245C c.3734T>G
p.F1245V c.3733T>G
p.F1245L c.3735C>G
p.F1245I c.3733T>A
p.112501 c.3749T>C
p.R12750 c.3824G>A
Table 3. EGFR Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.L747_T751>S c 2240 _2251de112 .
p.L8610 c.2582T>A
p.L747_E749de1 c.2239 _2247del9 p.E746_5752>D c 2238 _2255de118 .
p.E746_A750de1 c 2235 _2249de115 .
p.L858R c.2573T>G
p.E746_A750de1 c.2236 _2250de115 p.R776C c.2326C>T
p.H835L c.2504A>T
p.G719A c.2156G>C

p.T790M c.2369C>T
p.S7681 c.2303G>T
p.V769L c.2305G>T
p.G7195 c.2155G>A
p.G719C c.2155G>T
p.L747_T751de1 c 2239 _2253de115 .
p.L747_5752de1 c 2239 _2256de118 .
p.S752_1759de1 c 2254 _2277de124 .
p.P753S c.2257C>T
p.L858M c.2572C>A
p.E746_5752>A c 2237 _2254de118 .
p.L747_T751de1 c 2240 _2254de115 .
p.L747_P753>S c 2240 _2257de118 .
p.E709V c.2126A>T
p.17155 c.2144T>G
p.S720F c.2159C>T
p.L861R c.2582T>G
p.V769_D7701nsASV c 2307 _2308ins9 .
p.H773_V774insH c 2319 _2320insCAC
.
p.D770_N771insG c 2310 _2311insGGT
.
p.V769_D7701nsCV c 2307 _2308insTGCGTG
.
p.H773_V7741nsPH c 2319 _2320insCCCCAC
.
p.H773_V7741nsNPH c 2319 _2320ins9 .
p.L747_A750>P c.2239 _2248TTAAGAGAAG>C
p.L747_T751>P c 2239 _2251>C
.
p.E746_5752>V c 2237 _2255>T
.
p.E746_5752>1 c 2235 _2255>AAT
.
p.E746_T751>V c.2237 _2252>T

p.L747_P753>Q c 2239 _2258>CA
.
p.H773>NPY c.2317_2317C>AACCCCT
p.V774_C7751nsHV c.2322_2322G>CCACGTG
p.L747_5752>Q c.2239_2256>CAA
p.E746_T751>I c 2229 _2252>AATTAAGA
.
p.T751_1759>S c 2252 _2275>G
.
p.E746_A750>RP c.2236_2248>AGAC
p.E746_T751>VA c.2237_2253>TTGCT
p.L747_T751>Q c.2238_2252>GCA
p.L747_T751>Q c 2239 _2252>CA
.
p.L747_5752>QH c.2238_2255>GCAACA
p.L747_A750>P c 2238 _2248>GC
.
p.1744_K7451nsKIPVAI c 2231 _2232ins18 .
p.D761_E7621nsEAFQ c 2283 _2284ins12 .
p.A767_57681nsTLA c 2302 _2303ins9 .
p.V769_D7701nsASV c 2308 _2309ins9 .
p.D770>GY c 2308 _2309insGTT
.
p.E709H c 2125 _2127GAA>CAT
.
p.L858R c 2573 _2574TG>GT
.
p.A859T c.2575G>A
p.E746_T751>A c.2237_2251de115 p.Y727C c.2180A>G
p.V851I c.2551G>A
p.E746_T751de1 c.2236_2253de118 p.D770_N771>AGG c.2309 _2312ACAA>CTGGIGG
p.G857R c.2569G>A
p.L858R c.2573.T>G
p.E746_A750de1 c.2235 _2249de115 p.E746_A750>QP c.2236_2248>CAAC
p.G810D c.2429G>A
p.E709K c.2125G>A
p.D770_N771insN c.2310_23111nsAAC
p.D770_N771insG c.2310_23111nsGGC
p.H773L c.2318A>T
p.V774M c.2320G>A
p.G779F c.2335 2336GG>TT
p.A871G c.2612C>G
p.E709G c.2126A>G
p.L8610 c.25821>A
p.L730F c.2188C>T
p.P733L c.2198C>T
p.G735S c.2203G>A
p.V742A c.2225T>C
p.E746K c.2236G>A
p.1751I c.2252C>T
p.S752Y c.2255C>A
p.H850N c.2548C>A
p.D761N c.2281G>A
p.S784F c.2351C>T
p.L792P c.2375T>C
p.L798F c.2392C>T
p.G8105 c.2428G>A
p.N826S c.2477A>G
p.1847I c.2540C>T
p.V851A c.2552T>C
p.I8531 c.2558T>C

p.A864T c.2590G>A
p.E866K c.2596G>A
p.G873E c.2618G>A
p.E746_P753>LS c 2236 _2257>CTCT
.
p.V819V c.2457G>A
p.Y764Y c.2292C>T
p.L833V c.2497T>G
p.V769M c.2305G>A
p.L838V c.2512C>G
p.E709A c.2126A>C
p.D770_N7711nsSVD c 2311 _2312ins9 .
p.A839T c.2515G>A
p.H773R c.2318A>G
p.P772P c.2316C>T
p.E746_T751>A c 2235 _2251>AG
.
p.E746_A750>IP c 2235 _2248>AATTC
.
p.E746_1751>I c 2235 _2252>AAT
.
p.E746_T751>IP c 2235 _2251>AATTC
.
p.L858R c 2572 _2573CT>AG
.
p.N771_P772>SVDNR c.2312 _2315ACCC>GCGTGGACAACCG
p.D770_P772>ASVDNR c.2308_2315GACAACCC>CCAGCGTGGATAACCG
p.S752_1759de1 c 2253 _2276de124 .
p.E746_A750>QP c 2236 _2248>CAAC
.
p.V769_D7701nsASV c.2309 _2310AC>CCAGCGTGGAT
p.M766_A767insAl c.2298 _2299insGCCATA
p.G724S c.2170G>A
p.D770N c.2308G>A
p.1783I c.2348C>T

p.G863D c.2588G>A
p.V897I c.2689G>A
p.K745R c.2234A>G
p.P741L c.2222C>T
p.E734K c.2200G>A
p.E746de1 c.2234_2236delAGG
p.E746_T751>VP c.2237_2251>TTC
p.0787R c.2360A>G
p.V834L c.2500G>T
p.A755A c.2265C>T
p.G719D c.2156G>A
p.E746_5752>V c.2237_2256>TC
p.E746_P753>VS c 2237 _2257>TCT
.
p.E746_A750>DP c.2238_2249>TCC
p.V769_D7701nsGSV c.2308_23091ns9 p.V769_D7701nsGVV c.2308_23091ns9 p.N771>GF c.2311_2312AA>GGGIT
p.V774_C7751nsHV c.2321 _2322insCCACGT
p.G719C c 2154 _2155GG>TT
.
p.L747_R748>FP c.2241 _2244AAGA>CCCG
p.E872 c.2614G>T
p.G873G c.2619A>T
p.P753P c.2259G>A
p.G719fs*29 c.2156deIG
p.L747_K754>ST c.2240_2261>CGAC
p.S768_V7691nsVAS c.2303_23041ns9 p.V769_D7701nsDNV c.2307_23081ns9 p.D770_N7711nsAPW c.2310 _2311ins9 p.N771_P772insN c 2313 _2314insAAC
.
p.G796S c.2386G>A
p.E804G c.2411A>G
p.R841K c.2522G>A
p.V834M c.2500G>A
p.D761Y c.2281G>T
p.R776H c.2327G>A
p.L778L c.2334G>T
p.G779C c.2335G>T
p.P848L c.2543C>T
p.L747_T751>P c 2238 _2251>GC
.
p.T751_I759>S c 2251 _2277>TCT
.
p.N771>TH c 2311 _2312insCAC
.
p.H773_V7741nsPH c 2318 _2319insCCCCCA
.
p.V774_C7751nsHV c 2322 _2323insCACGTG
.
p.L862P c.2585T>C
p.S784Y c.2351C>A
p.F795S c.2384T>C
p.F795S c.2384T>C
p.Y813C c.2438A>G
p.Y801H c.2401T>C
p.C775Y c.2324G>A
p.D770_N7711nsDG c.2308 _2309insACGGCG
p.1751_1759>REA c.2252 _2277>GAGAAGCG
p.L7770 c.2330T>A
p.G7215 c.2161G>A
p.G721D c.2162G>A
p.K754K c.2262A>G

p.E746_1751>Q c.2236_2253>CAA
p.L747_T751de1 c.2238_2252de115 p.L747_T751>A c.2239_2253>GCT
p.C818Y c.2453G>A
p.I759N c.2276T>A
p.T751_E758de1 c.2250_2273de124 p.L747P c.2239_224011>CC
p.L858K c 2572 _2573CT>AA
.
p.P753_1759de1 c.2257_2277de121 p.T751_I759>N c 2252 _2277>AT
.
p.G863G c.2589T>G
p.N771>SH c.2311_23121nsGTC
p.D770fs*61 c.2309_23101ns14 p.E829E c.2487G>A
p.R831C c.2491C>T
p.R831C c.2491C>T
p.L861V c.2581C>G
p.E746_T751de1 c.2235_2252de118 p.L747_K754de1 c.2239 _2262de124 p.L838P c.2513T>C
p.K757>NK c.2270 _2271insCAA
p.G779S c.2335G>A
p.V774L c.2320G>T
p.L815L c.2445C>T
p.E758D c.2274A>C
p.K875R c.2624A>G
p.A864E c.2591C>A
p.Y869C c.2606A>G

p.K745_E749de1 c.2233_2247de115 p.F723F c.2169C>T
p.L858L c.2572C>T
p.A859fs*38 c.2575_2576insG
p.N756S c.2267A>G
p.V845A c.2534T>C
p.F856S c.2567T>C
p.G874S c.2620G>A
p.A750_E758de1 c.2247_2273de127 p.A750_E758>P c.2248_2273>CC
p.L747_5752>Q c.2238_2256>GCAA
p.A859_L883>V c.2576_2647de172 p.1744_K7451nsKIPVAI c.2232_22331ns18 p.K745_E7461nsVPVAIK c.2236_22371ns18 p.A767V c.2300C>T
p.N842D c.2524A>G
p.A743T c.2227G>A
p.L747S c.2240T>C
p.K860I c.2579A>T
p.A750_K754de1 c.2246_2260de115 p.D770_N7711nsMATP c.2311_23121ns12 p.A763_Y7641nsFQEA c.2290_22911ns12 p.D761G c.2282A>G
p.V786M c.2356G>A
p.G796A c.2387G>C
p.K728 c.2182A>T
p.R832C c.2494C>T
p.G721A c.2162G>C

p.I744V c.2230A>G
p.S784P c.2350T>C
p.R832L c.2495G>T
p.V802F c.2404G>T
p.E746_E749de1 c.2235_2246de112 p.T854A c.2560A>G
p.E884K c.2650G>A
p.F7125 c.2135T>C
p.I744M c.2232C>G
p.V765M c.2293G>A
p.R836C c.2506C>T
p.A871T c.2611G>A
p.D855G c.2564A>G
p.E868G c.2603A>G
p.L798H c.2393T>A
p.K806E c.2416A>G
p.L814P c.2441T>C
p.E746_A750>VP c.2237_2250>TCCCT
p.V769_D7701nsMASVD c.2307_23081ns15 p.F723S c.2168T>C
p.T785N c.2354C>A
p.V845M c.2533G>A
p.M766T c.2297T>C
p.S752P c.2254T>C
p.17251 c.2175G>A
p.D855N c.2563G>A
p.L8580 c.2573T>A
p.H870R c.2609A>G

p.F712L c.2134T>C
p.I8211 c.2462T>C
p.V834A c.2501T>C
p.L718P c.2153T>C
p.D770_N7711nsNPH c.2310_23111nsAACCCCCAC
p.D770_N7711nsGL c.2310_23111nsGGGITA
p.D770_N7711nsSVD c.2311_23121nsGCGTGGACA
p.P772_H7731nsTHP c.2315 _2316insGACACACCC
p.S720T c.2158T>A
p.E746V c 2237 _2238AA>TT
.
p.E746_P753>VQ c 2237 _2258>TTCA
.
p.E709_T710>D c 2127 _2129delAAC
.
p.E746_1751>IP c.2236_2253>ATTCCT
p.L747_T751>Q c 2239 _2253>CAA
.
p.H773_V7741nsGNPH c 2320 _2321ins12 .
p.I7321 c.2195T>C
p.N756Y c.2266A>T
p.L844P c.2531T>C
p.17401 c.2219T>C
p.E746_T751>VP c 2237 _2253>TTCCT
.
p.W731L c.2192G>T
p.E7340 c.2200G>C
p.T785A c.2353A>G
p.C797Y c.2390G>A
p.R831H c.2492G>A
p.N771>GY c.2311 _2311A>GGIT
p.P733S c.2197C>T
p.R748I c.2243G>T

p.0849R c.2546A>G
p.E746_T751>VA c 2237 _2251>TGG
.
p.E868D c.2604A>T
p.S7205 c.2160C>A
p.T725A c.2173A>G
p.R836S c.2506C>A
p.I7441 c.2232C>A
p.E866G c.2597A>G
p.I8531 c.2559C>T
p.K708E c.2122A>G
p.G824G c.2472C>A
p.F712F c.2136C>T
p.Y827Y c.2481C>T
p.T725M c.2174C>T
p.T725M c.2174C>T
p.K852N c.2556G>T
p.A722V c.2165C>T
p.E711K c.2131G>A
p.1785I c.2354C>T
p.D800N c.2398G>A
p.E872G c.2615A>G
p.E829K c.2485G>A
p.E829K c.2485G>A
p.H870Y c.2608C>T
p.H870Y c.2608C>T
p.D770_N7711nsSVD c 2310 _2311ins9 .
p.S768_V769>IL c.2303 _2305GCG>TCT
p.D770_N7711nsGD c.2310 _2311insGGGGAC

p.E709_T710>A c.2126_2128delAAA
p.E746_5752>V c.2235_2255>GGT
p.I744_A750>VK c.2230_2249>GTCAA
p.L747_K754>N c.2239_2264>GCCAA
p.1740_P7411nsPVAIKI c.2219_22201ns18 p.R836R c.2508C>T
p.V843I c.2527G>A
p.K754R c.2261A>G
p.A840T c.2518G>A
p.K754E c.2260A>G
p.A859D c.2576C>A
p.Y801C c.2402A>G
p.I7441 c.2231T>C
p.18541 c.2561C>T
p.G863S c.2587G>A
p.H850R c.2549A>G
p.K754A c.2260_2261AA>GC
p.D807N c.2419G>A
p.S720P c.2158T>C
p.K757M c.2270A>T
p.L8620 c.2585T>A
p.T751_1759>N c.2252_2276>A
p.P772R c.2315C>G
p.A839V c.2516C>T
p.K716R c.2147A>G
p.H773_V7741ns0 c.2319_23201nsCAG
p.E711V c.2132_2133AA>T1 p.T710A c.2128A>G

p.K714N c.2142G>C
p.V717A c.2150T>C
p.G729E c.2186G>A
p.I744_E749>LKR c 2230 _2247>CTTAAGAGA
.
p.E746_T751>L c 2236 _2253>CTA
.
p.E746_5752>1 c 2236 _2255>AT
.
p.E746_5752de1 c 2236 _2256deI21 .
p.E746_5752>1 c 2236 _2256>ATC
.
p.E746_P753>IS c 2236 _2259>ATCTCG
.
p.E746_T751>V c 2237 _2253>TA
.
p.E746_T751>V c 2237 _2253>TC
.
p.L747_A750>P c 2239 _2250>CCA
.
p.L747_5752>QH c.2239 _2256>CAACAT
p.L747_P753>S c 2239 _2257>T
.
p.R748K c.2243G>A
p.E749G c.2246A>G
p.T751_I759>S c 2251 _2277>TCC
.
p.P772_H7731nsHV c.2316 _2317insCACGTG
p.G779D c.2336G>A
p.V802A c.2405T>C
p.L833W c.2498T>G
p.D837G c.2510A>G
p.L844V c.2530C>G
p.T751_1759de1 c.2252 _2275de124 p.V765G c.2294T>G
p.G796D c.2387G>A
p.R836H c.2507G>A
p.K757R c.2270A>G

p.E872K c.2614G>A
p.L858L c.2574G>A
p.17805 c.2339T>G
p.T785P c.2353A>C
p.Y801fs*1 c.2402_2403insG
p.L858R c.2573 _2574TG>GA
p.N771_P7721nsRH c.2311_23121nsACCGGC
p.H850Y c.2548C>T
p.E868K c.2602G>A
p.17801 c.2339T>C
p.E866D c.2598G>T
p.L833F c.2499G>T
p.A864V c.2591C>T
p.K745_A750de1 c.2232_2249de118 p.P794H c.2381C>A
p.E804K c.2410G>A
p.G857E c.2570G>A
Table 4. KIT Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.(550_592)ins7 c.(1648_1774)ins21 p.C443Y c.1328G>A
p.P456S c.1366C>T
p.L462L c.1384C>T
p.P468P c.1404G>A
p.F469L c.1405T>C
p.L472L c.1416A>G

p.S4761 c.1427G>T
p.D479fs*2 c.1434_1462de129 p.S480fs*47 c.1439deIC
p.N486D c.1456A>G
p.V489I c.1465G>A
p.V489A c.1466T>C
p.E490G c.1469A>G
p.E490_F504>DHIVVSLTF c.1470_1512>CCACATCGTTGTAAGCCITACATTC
p.N495I c.1484A>T
p.N495I c.1484A>T
p.D496V c.1487A>T
p.V5OM c.148G>A
p.V497V c.1491G>A
p.G498D c.1493G>A
p.K499K c.1497G>A
p.A502_Y5031nsSA c.1504_1505insCTICTG
p.A502_Y5031nsSA c.1505_1506insTTCTGC
p.Y503_F5041nsSA c.1507_1508insCTGCCT
p.A502_Y5031nsFA c.1507_1508insTTGCCT
p.Y503_F5041nsAY c.1509_1510insGCCTAT
p.F504L c.1510T>C
p.N505H c.1513A>C
p.F506L c.1516T>C
p.F506_A5071nsAYFNF c 1518 _1519ins15 .
p.F508_K5091nsNFAF c.1524 _1525ins12 p.K5091 c.1526A>T
p.G510del c.1528 _1530deIGGT
p.N512D c.1534A>G

p.V530I c.1588G>A
p.I531_V5321nsGF c.1593_15941nsGGGITC
p.M541L c.1621A>C
p.K546K c.1638A>G
p.0549_V555>I c 1645 _1663>A
.
p.K550_P551de1 c.1648_1653delAAACCC
p.K550_E554de1 c.1648_1662de115 p.K550_V555de1 c.1648_1665de118 p.K550_V555de1 c.1648_1665de118 p.K550_0556>I1 c 1648 _1668>ATTATT
.
p.K550_W557de1 c.1648_1671de124 p.K550fs*6 c 1648 _1672de125 .
p.K550_K558de1 c.1648_1674de127 p.K550_V559de1 c.1648_1677de130 p.K550_V555>I c.1649_1663de115 p.K550R c.1649A>G
p.K550I c.1649A>T
p.K550_V555>KTL c.1650_1663>AACCC
p.P551_K558de1 c.1650_1673de124 p.K550N c.1650A>C
p.P551de1 c 1651 _1653delCCC
.
p.P551_E554de1 c.1651_1662de112 p.P551_V555de1 c.1651_1665de115 p.P551_0556de1 c.1651 _1668de118 p.P551T c.1651C>A
p.P5515 c.1651C>T
p.P551_M552>L c.1652_1654delCCA
p.P551_E554>H c.1652_1662>AY

p.P551_V555>L c.1652_1663de112 p.P551_V559de1>L c.1652_1678de127 p.P551L c.1652C>T
p.M552_Y553de1 c.1653_1658deICATGTA
p.M552_0556> c 1653 _1667>TCT
.
p.M552_W557de1 c.1653_1670de118 p.M552_Y553de1 c.1654_1659delATGTAT
p.M552_E554de1 c.1654_1662de19 p.M552_V555de1 c.1654_1665de112 p.M552_0556de1 c 1654 _1668de115 .
p.M552_W557de1 c.1654_1671de118 p.M552_K558de1 c.1654_1674de121 p.M552_D572de1 c.1654_1716de163 p.M552L c.1654A>C
p.M552L c.1654A>C
p.M552_Y553>N c.1655_1657deITGT
p.M552_E554>K c.1655_1660deITGTATG
p.M552_V555>1 c.1655_1663de19 p.M552_0556>K c 1655 _1666de112 .
p.M552_W557>R c.1655_1669de115 p.M552_W557de1 c.1655_1672de118 p.M552_K558>T c.1655_1674>CN
p.M552_E561>K c.1655_1681de127 p.M552_T574>TESA c.1655 _1720>CAGAATCAG
p.M552K c.1655T>A
p.M552T c.1655T>C
p.Y553_W557de1 c.1656_1670de115 p.Y553_K558> c.1656_1673de118 p.Y553V c 1657 _1658TA>GT
.
p.Y553_0556de1 c 1657 _1668de112 .
p.Y553_W557de1 c.1657_1671de115 p.Y553_K558de1 c.1657_1674de118 p.Y553_V559>E c.1657_1677>GAA
p.Y553_V559de1 c.1657_1677de121 p.Y553N c.1657T>A
p.Y553_T574>S c.1658_1720de163 p.E554_K558de1 c.1660_1674de115 p.E554_E562de1 c.1660_1686de127 p.E554_N564de1 c.1660_1692de133 p.E554_1571de1 c.1660_1713de154 p.E554_D572de1 c.1660_1716de157 p.E554K c.1660G>A
p.E554K c.1660G>A
p.E554_K558de1 c.1661_1675de115 p.E554G c.1661A>G
p.V555_E562de1 c.1662_1685de124 p.E554D c.1662A>T
p.V555_0556de1 c.1663 _1668delGTACAG
p.V555_K558de1 c.1663_1674de112 p.V555_V559de1 c.1663_1677de115 p.V555_V560de1 c.1663_1680de118 p.V555_1563de1 c.1663_1689de127 p.V555_G565de1 c.1663_1695de133 p.V555_Y570de1 c.1663_1710de148 p.V555_1571de1 c.1663_1713de151 p.V555_P573de1 c.1663_1719de157 p.V555I c.1663G>A
p.V555_N566>D c.1664_1696de133 p.0556_V559de1 c.1665_1676de112 p.V555_V560>V c.1665_1679de115 p.0556_N566>SNNLQLY c.1665 _1696>TTCCAACAACCITCCACTGT
p.0556_D572de1 c.1665_1716>T
p.0556_W557de1 c.1666_1671deICAGIGG
p.0556_V559de1 c.1666_1677de112 p.0556_V560>F c.1666_1678>T
p.0556_V560>TTF c.1666_1680>ACAACCITC
p.0556_V560de1 c.1666_1680de115 p.0556_E561>HH c.1666_1683>CATCAT
p.0556_E561de1 c.1666_1683de118 p.0556_D572>PS c.1666_1716>CCATCC
p.0556_P573de1 c.1666_1719de154 p.0556_T574de1 c 1666 _1722de157 .
p.0556_L576de1 c.1666_1728de163 p.0556_W557>R c.1667_1669delAGT
p.W557_K558de1 c.1667_1672delAGIGGA
p.0556_K558>R c.1667_1673AGIGGAA>G
p.W557_E561de1 c.1667_1681de115 p.0556R c.1667A>G
p.W557_K558de1 c.1668_1673delGTGGAA
p.0556_K558>HPCR
c.1668_1673GTGGAA>CCCCTGCAG
p.0556_K558>H c.1668_1674GTGGAAG>Y
p.0556_V559>H c.1668_1676de19 p.0556_V559>HT
c.1668_1677GTGGAAGGIT>TACT
p.0556_V560>HNLQLY c.1668 _1679>CAACCITCCACTGTA

p.0556_V560>H c 1668 _1679de112 .
p.W557_1571de1 c.1668_1712de145 p.0556_D572>H c 1668 _1715de148 .
p.W557_0575de1 c 1668 _1724de157 .
p.W557de1 c 1669 _1671deITGG
.
p.W557_K558>E c.1669_1672TGGA>G
p.W557_K558de1 c.1669_1674deITGGAAG
p.W557_K558>S c.1669_1674TGGAAG>C
p.W557_V559>I c.1669_1675TGGAAGG>A
p.W557_V559de1 c.1669_1677de19 p.W557_V560de1 c.1669_1680de112 p.W557_E561de1 c.1669_1683de115 p.W557_E562de1 c.1669_1686de118 p.W557_0575de1 c 1669 _1725de157 .
p.W557R c.1669T>A
p.W557R c.1669T>C
p.W557G c.1669T>G
p.W557_K558>SS c.1670_1673GGAA>CTTC
p.W557_K558>FP c.1670_1674GGAAG>TTCCT
p.W557_V559>F c.1670_1675deIGGAAGG
p.W557_V560>F c.1670_1678de19 p.W557_P573>S c.1670_1717de148 p.W557S c.1670G>C
p.W557_K558>CT c.1671_1673GAA>CAC
p.W557_K558>CP c.1671_1673GAA>TCC
p.W557_K558>C c.1671_1674GAAG>C
p.W557_V559>C c.1671_1676delGAAGGT
p.W557_V560>C c.1671_1679de19 p.W557 c.1671G>A
p.W557C c.1671G>T
p.K558_V559>SS c. 1672 _1676AAGGT>TCTTC
p.K558_V559de1 c.1672_1677delAAGGIT
p.K558_V560de1 c.1672_1680de19 p.K558_E562de1 c.1672_1686de115 p.K558_N564de1 c.1672_1692de121 p.K558_G565de1 c.1672_1695de124 p.K558_D572de1 c.1672_1716de145 p.K558_0575de1 c 1672 _1725de154 .
p.K558E c.1672A>G
p.K558* c.1672A>T
p.K558>NP c 1673 _1674insTCC
.
p.K558_V560>1 c.1673_1678delAGGITG
p.K558_V560>M c. 1673 _1680AGGITGIT>TG
p.K558_E562de1 c.1673_1687de115 p.K558_G565>R c.1673_1693de121 p.K558R c.1673A>G
p.K558>NP c 1674 _1674G>TCCT
.
p.K558_V559>N c.1674_1676deIGGT
p.K558_V560>N c.1674_1679deIGGITGT
p.K558_Y570>N c.1674_1709de136 p.K558_1_576>NV c 1674 _1726>CG
.
p.K558K c.1674G>A
p.K558N c.1674G>C
p.K558N c.1674G>Y
p.V559de1 c 1675 _1677delGTT
.
p.V559K c.1675 _1677GTT>AAG

p.V559_V560de1 c.1675_1680delGTTGIT
p.V559_E561de1 c.1675_1683de19 p.V559_G565de1 c 1675 _1695deI21 .
p.V559_1571de1 c 1675 _1713de139 .
p.V559_L576de1 c.1675_1728de154 p.V559I c.1675G>A
p.V559_E561de1 c.1676_1684de19 p.V559_E562de1 c.1676_1687de112 p.V559_P573>A c 1676 _1717de142 .
p.V559D c.1676T>A
p.V559A c.1676T>C
p.V559G c.1676T>G
p.V560de1 c 1678 _1680delGTT
.
p.V560_L576de1 c.1678_1728de151 p.V560E c 1679 _1680TT>AG
.
p.V560E c 1679 _1680TT>AR
.
p.V560de1 c 1679 _1681deITTG
.
p.V560_1571de1 c 1679 _1714de136 .
p.V560D c.1679T>A
p.V560A c.1679T>C
p.V560G c.1679T>G
p.E561de1 c 1680 _1682deITGA
.
p.V560V c.1680T>G
p.E561de1 c.1681 _1683delGAG
p.E561_P577de1 c.1681 _1731deI51 p.E561K c.1681G>A
p.E561G c.1682A>G
p.E561E c.1683G>A

p.E562_P573de1 c.1684_1719de136 p.E562K c.1684G>A
p.E562V c.1685A>T
p.E562_V569>D c.1686_1706de121 p.1563_D572de1 c.1687_1716de130 p.1563_L576de1 c.1687_1728de142 p.I563V c.1687A>G
p.N564_T574de1 c.1690_1722de133 p.N564_L576de1 c.1690_1728de139 p.N564_P577de1 c.1690_1731de142 p.N564_Y578de1 c.1690_1734de145 p.N564H c.1690A>C
p.N564_P573>TS c.1691_1717>CCT
p.N564_P573>T c.1691_1717de127 p.N564S c.1691A>G
p.N564K c.1692T>G
p.G565R c.1693G>A
p.G565E c.1694G>A
p.G565V c.1694G>T
p.N566D c.1696A>G
p.N566S c.1697A>G
p.N567_L576>E c.1698_1728>CGAA
p.N566N c.1698C>T
p.N567_P573de1 c.1699_1719de121 p.N567H c.1699A>C
p.N567K c.1701T>A
p.Y568_T574de1 c.1702 _1722deI21 p.Y568D c.1702T>G

p.Y568_1376>CV c 1703 _1726>GTG
.
p.Y568S c.1703A>C
p.Y568C c.1703A>G
p.Y568Y c.1704T>C
p.V569_L576de1 c.1705_?del?
p.V569_D572de1 c.1705_1716de112 p.V569_0575de1 c.1705_1725de121 p.V569_L576de1 c.1705_1728de124 p.V569I c.1705G>A
p.Y570_1376delYIDPTQL c.1706_1726de121 p.V569_L576>G c 1706 _1727>G
.
p.V569A c.1706T>C
p.V569G c.1706T>G
p.Y570_L576de1 c.1708_1728de121 p.Y570D c.1708T>G
p.Y570* c.1710C>A
p.1571_L576de1 c.1711_1728de118 p.1571_N587de1 c.1712_1762de151 p.I571R c.1712T>G
p.I571M c.1713A>G
p.571_572>GE c.1714 _1715insGGGAAG
p.D572N c.1714G>A
p.D572Y c.1714G>T
p.D572A c.1715A>C
p.D572D c.1716C>T
p.P573L c.1718C>T
p.P573_T5741nsYlDP c.1719 _1720ins12 p.T574A c.1720A>G

p.T574_05751ns12 c.1721_17221ns36 p.1574I c.1721C>T
p.0575de1 c.1723_1725deICAA
p.0575_P577>T c.1723_1731CAACTTCCT>ACA
p.L576de1 c.1726_1728deIC11 p.L576F c.1726C>T
p.L576de1 c.1727_1729deI11C
p.L576P c.1727T>C
p.L576_P577insQL c.1728_17291nsCAACTT
p.P577_Y578de1 c.1729_1734delCCITAT
p.P577S c.1729C>T
p.P577_D579de1 c.1730_1738del9 p.P577H c.1730C>A
p.P577L c.1730C>T
p.D579de1 c.1735_1737delGAT
p.D579_H5801nsPTQLPYD c.1737_1738ins21 p.D579_H5801nsSYD c.1737_17381ns9 p.H580de1 c.1737_1739deITCA
p.H580Y c.1738C>T
p.H580_K5811nsHPYD c.1739_17401ns12 p.H580_K5811nsPYDH c.1740_17411ns12 p.H580_K5811nsPTQLPYDH c.1740_17411ns24 p.H580_K5811nsIDPTQLPYDH c.1740_17411ns30 p.H580_K5811nsYDH c.1740_17411ns9 p.K581R c.1742A>G
p.W582* c.1745G>A
p.W582* c.1746G>A
p.E583_F5841nsPYDHKWE c.1748_1749ins21 p.E583G c.1748A>G
p.F584L c.1750T>C
p.F584S c.1751T>C
p.F584_P5851nsLPYDHKWEF c.1752_17531ns27 p.F584_P5851ns13 c.1752_17531ns39 p.F584_P5851ns15 c.1752_17531ns45 p.P585_R5861nsYDHKWEFP c.1754_17551ns24 p.P585_R5861ns12 c.1754_17551ns36 p.P585_R5861nsLPYDHKWEFP c.1755_17561ns30 p.P585_R5861ns13 c.1755_17561ns39 p.P585_R5861ns14 c.1755_17561ns42 p.P585_R5861ns17 c.1755_17561ns51 p.P585P c.1755C>T
p.N587_R5881ns15 c.1761_17621ns45 p.N587N c.1761C>T
p.R588_L5891ns17 c.1764_17651ns51 p.S590N c.1769G>A
p.F591L c.1771T>C
p.F591_G5921ns21 c.1773_17741ns63 p.G592_K5931ns16 c.1774_17751ns48 13.? c.1774+3C>T
p.G592_K5931ns21 c.1775_17761ns63 p.1594I c.1781C>T
p.A599T c.1795G>A
p.P627L c.1880C>T
p.1632I c.1895C>T
p.E633G c.1898A>G
p.R634R c.1902G>A

p.E635G c.1904A>G
p.A636V c.1907C>T
p.L637F c.1909C>T
p.S639P c.1915T>C
p.K6420 c.1924A>C
p.K642E c.1924A>G
p.V643A c.1928T>C
p.S645N c.1934G>A
p.L647F c.1939C>T
p.L647P c.1940T>C
p.G648S c.1942G>A
p.N649_H650insN c.1947_19481nsAAT
p.I6531 c.1958T>C
p.V654A c.1961T>C
p.N655K c.1965T>G
p.G663V c.1988G>T
p.G664R c.1990G>A
p.1670E c.2008_2009AC>GA
p.1670I c.2009C>T
p.L682fs*1 c.2045de11 p.S692L c.2075C>T
p.E695K c.2083G>A
p.H697Y c.2089C>T
p.H697fs*28 c.2089deIC
p.R815_D816insVI c.2445_24461nsGTCATA
p.D816I c.2446_2447GA>AT
p.D816F c.2446_2447GA>TT
p.D816N c.2446G>A

p.D816H c.2446G>C
p.D816Y c.2446G>T
p.D816>GP c.2447_2448AC>GGCCA
p.D816>VVA c.2447_2448AC>TCGTTGCA
p.D816A c.2447A>C
p.D816G c.2447A>G
p.D816V c.2447A>T
p.D816E c.2448C>G
p.D816E c.2448C>G
p.I817V c.2449A>G
p.I8171 c.2450T>C
p.K818R c.2453A>G
p.K818K c.2454G>A
p.N819Y c.2455A>T
p.D820N c.2458G>A
p.D820H c.2458G>C
p.D820H c.2458G>C
p.D820Y c.2458G>T
p.D820Y c.2458G>T
p.D820A c.2459A>C
p.D820G c.2459A>G
p.D820V c.2459A>T
p.D820E c.2460T>A
p.D820E c.2460T>G
p.N822H c.2464A>C
p.N822Y c.2464A>T
p.N822Y c.2464A>T
p.N822S c.2465A>G

p.N822K c.2466T>A
p.N822N c.2466T>C
p.N822K c.2466T>G
p.N822K c.2466T>R
p.Y823N c.2467T>A
p.Y823D c.2467T>G
p.Y823C c.2468A>G
p.V825I c.2473G>A
p.V825A c.2474T>C
p.A829P c.2485G>C
p.A829V c.2486C>T
p.R830* c.2488C>T
p.R830* c.2488C>T
p.L831P c.2492T>C
p.V833L c.2497G>C
p.V833V c.2499G>T
p.E839K c.2515G>A
p.C844Y c.2531G>A
p.Y846H c.2536T>C
p.F848L c.2542T>C
p.E849* c.2545G>T
p.W853* c.2558G>A
p.S854P c.2560T>C
p.L859P c.2576T>C
p.L859L c.2577T>G
p.E861E c.2583G>A
p.L862L c.2586G>C

Table 5. KRAS Mutations That Are Covered Amino Acid Mutation Nucleotide Mutation p.V9V c.27T>C
p.A11P c.31G>C
p.A11V c.32C>T
p.G12F c.34_35GG>TT
p.G12C c.34_36GGT>TGC
p.G12L c.34_35GG>CT
p.G12L c.34_35GG>CT
p.G12V c 35 _36GT>TC
.
p.G12C c.34G>T
p.G125 c.34G>A
p.G12R c.34G>C
p.G12E c.35_36GT>AA
p.G12V c.35G>T
p.G12D c.35G>A
p.G12A c.35G>C
p.G12G c.36T>C
p.G13C c.37G>T
p.G135 c.37G>A
p.G13R c.37G>C
p.G13D c.38G>A
p.G13A c.38G>C
p.G13V c.38G>T
p.A18T c.52G>A
p.A18D c.53C>A
p.061K c.181C>A
p.061E c.181C>G

p.061P c.182A>C
p.061R c.182A>G
p.061L c.182A>T
p.061H c.183A>C
p.061H c.183A>T
p.D69fs*4 c.205deIG
p.G12fs*3 c.35deIG
p.G13V c.38_39GC>TT
p.V14I c.40G>A
p.061K c.180 _181TC>CA
Table 6. BRAF Mutations That Are Covered Amino Acid Mutation cDNA Nucleotide Mutation p.G3OD c.89G>A
p.M53T c.158T>C
p.S102F c.305C>T
p.S129L c.386C>T
p.R146W c.436C>T
p.11561 c.468C>T
p.R178* c.532C>T
p.A184T c.550G>A
p.Y198H c.592T>C
p.Q201H c.603G>T
p.K205Q c.613A>C
p.F247L c.741T>G
p.Q257H c.771G>T

p.G258V c.773G>T
p.H298Y c.892C>T
p.I300V c.898A>G
p.A305V c.914C>T
p.E309* c.925G>T
p.T310I c.929C>T
p.S323S c.969G>A
p.I326V c.976A>G
p.I326T c.977T>C
p.F357S c.1070T>C
p.G358G c.1074G>C
p.S364L c.1091C>T
p.S365L c.1094C>T
p.P367R c.1100C>G
p.S394* c.1181C>G
p.T401I c.1202C>T
p.P403fs*8 c.1208delC
p.A404fs*9 c.1208_1209insC
p.G421V c.1262G>T
p.G421G c.1263A>G
p.K439Q c.1315A>C
p.K439T c.1316A>C
p.T440P c.1318A>C
p.T440A c.1318A>G
p.T440T c.1320A>G
p.G442S c.1324G>A
p.R444W c.1330C>T

p.R444Q c.1331G>A
p.R444L c.1331G>T
p.R444R c.1332G>A
p.R444R c.1332G>T
p.S447S c.1341T>C
p.W450* c.1349G>A
p.W450L c.1349G>T
p.P453T c.1357C>A
p.P453P c.1359T>C
p.G455R c.1363G>A
p.G455E c.1364G>A
p.Q456* c.1366C>T
p.Q456R c.1367A>G
p.Q456Q c.1368G>A
p.I457T c.1370T>C
p.V459L c.1375G>C
p.V459A c.1376T>C
p.V459V c.1377G>A
p.G460* c.1378G>T
p.G460G c.1380A>G
p.R462G c.1384A>G
p.R462K c.1385G>A
p.R462I c.1385G>T
p.R462R c.1386A>G
p.I463V c.1387A>G
p.I463S c.1388T>G
p.1463I c.1389T>C

p.G464R c.1390G>A
p.G464R c.1390G>C
p.G464E c.1391G>A
p.G464V c.1391G>T
p.S465S c.1395T>C
p.G466R c.1396G>A
p.G466R c.1396G>C
p.G466E c.1397G>A
p.G466A c.1397G>C
p.G466V c.1397G>T
p.G466G c.1398A>G
p.S467P c.1399T>C
p.S467L c.1400C>T
p.F468L c.1402T>C
p.F468S c.1403T>C
p.F468C c.1403T>G
p.F468F c.1404T>C
p.G469R c.1405G>A
p.G469R c.1405G>C
p.G469>? c.1405 1406GG>CT
p.G469S c.1405 1406GG>TC
p.G469L c.1405 1406GG>TT
p.G469S c.1405 1407GGA>AGC
p.G469S c.1405 1407GGA>AGT
p.G469E c.1406G>A
p.G469A c.1406G>C
p.G469V c.1406G>T

p.G469G c.1407A>G
p.V471I c.1411G>A
p.V471F c.1411G>T
p.V471A c.1412T>C
p.Y472S c.1415A>C
p.Y472C c.1415A>G
p.K475R c.1424A>G
p.K475M c.1424A>T
p.K475K c.1425G>A
p.D479Y c.1435G>T
p.L485L c.1453T>C
p.L485S c.1454T>C
p.L485 P490>Y c 1454 1469>A
. _ p.L485F c.1455G>T
p.N486 P490del c.1457_1471de115 p.V487V c.1461G>A
p.L505H c.1514T>A
p.R509* c.1525C>T
p.L514P c.1541T>C
p.W531C c.1593G>T
p.L537S c.1610T>C
p.H539P c.1616A>C
p.H542Y c.1624C>T
p.K570K c.1710G>A
p.H574N c.1720C>A
p.H574Q c.1722C>A
p.N581S c.1742A>G

p.N581I c.1742A>T
p.I582M c.1746A>G
p.F583S c.1748T>C
p.F583F c.1749T>C
p.L584F c.1750C>T
p.L584P c.1751T>C
p.L584L c.1752T>C
p.H585H c.1755T>C
p.E586K c.1756G>A
p.E586E c.1758A>G
p.D587N c.1759G>A
p.D587A c.1760A>C
p.D587G c.1760A>G
p.D587E c.1761C>A
p.D587E c.1761C>G
p.L588P c.1763T>C
p.L588R c.1763T>G
p.L588L c.1764C>T
p.T589A c.1765A>G
p.T589I c.1766C>T
p.T589T c.1767A>G
p.V590I c.1768G>A
p.V590A c.1769T>C
p.V590fs*3 c.1769delT
p.V590V c.1770A>G
p.K591E c.1771A>G
p.K591R c.1772A>G

p.I592V c.1774A>G
p.I592M c.1776A>G
p.1592I c.1776A>T
p.G593S c.1777G>A
p.G593C c.1777G>T
p.G593D c.1778G>A
p.D594N c.1779_1780TG>GA
p.D594N c.1780G>A
p.D594H c.1780G>C
p.D594G c.1781A>G
p.D594V c.1781A>T
p.D594E c.1782T>A
p.D594D c.1782T>C
p.D594E c.1782T>G
p.F595L c.1783T>C
p.F595S c.1784T>C
p.F595L c.1785T>A
p.F595F c.1785T>C
p.F595L c.1785T>G
p.G596R c.1786G>C
p.G596fs*2 c.1786de1G
p.G596D c.1787G>A
p.G596G c.1788T>C
p.L597V c.1789C>G
p.L597S c.1789_1790CT>TC
p.L597Q c.1790T>A
p.L597P c.1790T>C

p.L597R c.1790T>G
p.L597L c.1791A>G
p.A598T c.1792G>A
p.A598V c.1793C>T
p.A598A c.1794T>A
p.A598 T599insV c.1794 1795insGTT
p.T599de1 c.1794 1796delTAC
p.T599I c.1796C>T
p. T599 V600insT c.1796 1797insTAC
p.T599 V600>IAL c.1796 1798CAG>TAGCTT
p. T599 R603>I c.1796 1809>TC
p.T599T c.1797A>B
p.T599T c.1797A>G
p.T599T c.1797A>T
p. T599 V600insTT c.1797 1797A>TACTACG
p.T599 V600insTT c.1797 1798ins?
p. T599 V600insT c.1797 1798insACA
p.T599 V600insDFGLAT c.1798 1799ins18 p.V600R c.1797 1799AGT>GAG
p.V600M c.1798G>A
p.V600L c.1798G>C
p.V600L c.1798G>T
p.V600>YM c.1798 1798G>TACA
p.V600K c.1798 1799GT>AA
p.V600R c.1798 1799GT>AG
p.V600Q c.1798 1799GT>CA
p.V600E c.1799T>A

p.V600A c.1799T>C
p.V600G c.1799T>G
p.V600E c.1799_1800TG>AA
p.V600D c.1799_1800TG>AC
p.V600D c.1799_1800TG>AT
p.V600fs*11 c.1799_1800de1TG
p.V600 K601>E c. 1799 _1801delTGA
p.V600 S602>DT c. 1799 _1804TGAAAT>ATA
p.V600 S605>D c. 1799 _1814>A
p.V600 S605>DV c. 1799 1814>ATGT
_ p.V600 S605>EK c. 1799 _1815>AAAAG
p.V600V c.1800G>A
p.V600? c.(1798-1800)?
p.K601E c.1801A>G
p.K6Oldel c.1801_1803de1AAA
p.K601R c.1802A>G
p.K601I c.1802A>T
p.K601N c.1803A>C
p.K601K c.1803A>G
p.K601N c.1803A>T
p.S602S c.1806T>G
p.R603R c.1807C>A
p.R603* c.1807C>T
p.R603L c.1808G>T
p.R603R c.1809A>G
p.W604de1 c.1808_1810delGAT
p.W604R c.1810T>A

p.W604G c.1810T>G
p.W604* c.1811G>A
p.W604* c.1812G>A
p.S605G c.1813A>G
p.S605F c.1813_1814AG>TT
p.S605N c.1814G>A
p.S605R c.1815T>A
p.G606R c.1816G>A
p.G606S c.1816_1818GGG>AGT
p.G606E c.1817G>A
p.G606A c.1817G>C
p.G606V c.1817G>T
p.G606G c.1818G>A
p.S607P c.1819T>C
p.H608R c.1823A>G
p.H608H c.1824T>C
p.Q609R c.1826A>G
p.Q609Q c.1827G>A
p.F610L c.1828T>C
p.F610S c.1829T>C
p.F610F c.1830T>C
p.E611G c.1832A>G
p.E611D c.1833A>C
p.E611E c.1833A>G
p.Q612E c.1834C>G
p.Q612* c.1834C>T
p.S614P c.1840T>C

p.S614S c.1842T>C
p.G615R c.1843G>A
p.S616P c.1846T>C
p.S616F c.1847C>T
p.I617T c.1850T>C
p.L618L c.1852T>C
p.L618S c.1853T>C
p.L618W c.1853T>G
p.W619R c.1855T>C
p.Q636E c.1906C>G
p.Q636* c.1906C>T
p.Q636R c.1907A>G
p.S637P c.1909T>C
p.S637* c.1910C>G
p.S637L c.1910C>T
p.S657S c.1971A>G
p.R671Q c.2012G>A
p.R682W c.2044C>T
p.R682Q c.2045G>A
p.K698R c.2093A>G
p.A718V c.2153C>T
p.P731S c.2191C>T
p.P731P c.2193C>T

Table 7. ALK Capture Primers List for NGS Panel ¨ Genomic DNA
ALK Exon21 130-150 bases Seq. . Seq. Primer Sequence Pnmer Sequence ID ID
1 Left CCTCTTGTCTTCTCCTTTGCAC 21 Left CCTCTTGTCTTCTCCTTTGCAC
2 Right GGGCAGGCTCAAGAGTGA 22 Right AGCCAAGGGCAGGCTCAA
3 Left CCTCTTGTCTTCTCCTTTGCAC 23 Left CTCTTGTCTTCTCCTTTGCAC
4 Right AGGGCAGGCTCAAGAGTGA 24 Right AGGGCAGGCTCAAGAGTG
Left CCTCTTGTCTTCTCCTTTGCAC 25 Left CCTCTTGTCTTCTCCTTTGC
6 Right AAGGGCAGGCTCAAGAGTGA 26 Right GGGCAGGCTCAAGAGTGA
7 Left CCTCTTGTCTTCTCCTTTGCAC 27 Left CCTCTTGTCTTCTCCTTTGC
8 Right CAAGGGCAGGCTCAAGAGTGA 28 Right AGGGCAGGCTCAAGAGTGA
9 Left CTCTTGTCTTCTCCTTTGCAC 29 Left CTCTTGTCTTCTCCTTTGCAC
Right CAAGGGCAGGCTCAAGAGT 30 Right CAAGGGCAGGCTCAAGAGTG
11 Left CTCTTGTCTTCTCCTTTGCAC 31 Left CCTCTTGTCTTCTCCTTTGC

12 Right AAGGGCAGGCTCAAGAGTG 32 Right AAGGGCAGGCTCAAGAGTGA

13 Left CTCTTGTCTTCTCCTTTGCAC 33 Left CCTCTTGTCTTCTCCTTTGCAC

14 Right GGGCAGGCTCAAGAGTGA 34 Right AGCCAAGGGCAGGCTCAAGAGTGA
Left CTCTTGTCTTCTCCTTTGCAC 35 Left TCTTGTCTTCTCCTTTGCAC
16 Right AGGGCAGGCTCAAGAGTGA 36 Right CAAGGGCAGGCTCAAGAGT
17 Left CCTCTTGTCTTCTCCTTTGCAC 37 Left TCTTGTCTTCTCCTTTGCAC
18 Right CCAAGGGCAGGCTCAAGAGTGA 38 Right AAGGGCAGGCTCAAGAGTG
19 Left CTCTTGTCTTCTCCTTTGCAC 39 Left CTCTTGTCTTCTCCTTTGCAC
Right AAGGGCAGGCTCAAGAGTGA 40 Right CCAAGGGCAGGCTCAAGAGT
ALK Exon21 151-200 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
41 Left ACTCTGTCTCCTCTTGTCTTCTCCT 51 Left CTCTGTCTCCTCTTGTCTTCTCCT
42 Right CTGAGAACTGCAGCCTACAGAGT 52 Right CTGAGAACTGCAGCCTACAGAGT
43 Left CTCTGTCTCCTCTTGTCTTCTCCTT 53 Left TTGACTCTGTCTCCTCTTGTCTTCT
44 Right CTGAGAACTGCAGCCTACAGAGT 54 Right CTGAGAACTGCAGCCTACAGAG
45 Left TCTGTCTCCTCTTGTCTTCTCCTT 55 Left CTGTCTCCTCTTGTCTTCTCCTTT
46 Right CTGAGAACTGCAGCCTACAGAGT 56 Right CTGAGAACTGCAGCCTACAGAGT
47 Left TCTGTCTCCTCTTGTCTTCTCCTTT 57 Left CTCTGTCTCCTCTTGTCTTCTCCTT
48 Right CTGAGAACTGCAGCCTACAGAGT 58 Right AGAACTGCAGCCTACAGAGTCC
49 Left ACTCTGTCTCCTCTTGTCTTCTCCT 59 Left ACTCTGTCTCCTCTTGTCTTCTCCT
50 Right AGAACTGCAGCCTACAGAGTCC 60 Right CTGAGAACTGCAGCCTACAGAG

61 Left GTTTGACTCTGTCTCCTCTTGTCTT 71 Left CTCTGTCTCCTCTTGTCTTCTCCT
62 Right CTGAGAACTGCAGCCTACAGAG 72 Right AGAACTGCAGCCTACAGAGTCC
63 Left TCTGTCTCCTCTTGTCTTCTCCTT 73 Left CTGTTTGACTCTGTCTCCTCTTGTC
64 Right AGAACTGCAGCCTACAGAGTCC 74 Right CTGAGAACTGCAGCCTACAGAG
65 Left ACTCTGTCTCCTCTTGTCTTCTCCT 75 Left CTCTGTCTCCTCTTGTCTTCTCCTT
66 Right GAGAACTGCAGCCTACAGAGTCC 76 Right CTGAGAACTGCAGCCTACAGAG
67 Left TCTGTCTCCTCTTGTCTTCTCCTTT 77 Left CTGTCTCCTCTTGTCTTCTCCTTT
68 Right AGAACTGCAGCCTACAGAGTCC 78 Right AGAACTGCAGCCTACAGAGTCC
69 Left CTGTCTCCTCTTGTCTTCTCCTTTG 79 Left ACTCTGTCTCCTCTTGTCTTCTCC
70 Right CTGAGAACTGCAGCCTACAGAGT 80 Right AGAACTGCAGCCTACAGAGTCC
ALK Exon21 201-300 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
81 Left TGTTGAGGGTATTACTCCTGAGTGT _ 01 Left TGTTGAGGGTATTACTCCTGAGTGT
82 Right CTGAGAACTGCAGCCTACAGAGT _ 02 Right TGAGAACTGCAGCCTACAGAGT
83 Left TTGAGGGTATTACTCCTGAGTGTGT _ 03 Left TTGAGGGTATTACTCCTGAGTGTGT
84 Right CTGAGAACTGCAGCCTACAGAGT _ 04 Right TGAGAACTGCAGCCTACAGAGT
85 Left GTTGAGGGTATTACTCCTGAGTGTG _ 05 Left TGAGGGTATTACTCCTGAGTGTGT
86 Right AGAACTGCAGCCTACAGAGTCC t 06 Right CTGAGAACTGCAGCCTACAGAGT
87 Left TGTTGAGGGTATTACTCCTGAGTGT I_ 07 Left GTTGAGGGTATTACTCCTGAGTGTG
88 Right AGAACTGCAGCCTACAGAGTCC _ 08 Right GAGAACTGCAGCCTACAGAGTCC
89 Left TTGAGGGTATTACTCCTGAGTGTGT _ 09 Left TGTTGAGGGTATTACTCCTGAGTGT
90 Right AGAACTGCAGCCTACAGAGTCC _ 10 Right GAGAACTGCAGCCTACAGAGTCC
91 Left CTCTCGTGTTTGTCCACTAAATGT _ 11 Left TTGAGGGTATTACTCCTGAGTGTGT

92 Right CTGAGAACTGCAGCCTACAGAGT _ 12 Right GAGAACTGCAGCCTACAGAGTCC
93 Left TGAGGGTATTACTCCTGAGTGTGTAT _ 13 Left GTTGAGGGTATTACTCCTGAGTGTGT
94 Right CTGAGAACTGCAGCCTACAGAGT _ 14 Right CTGAGAACTGCAGCCTACAGAGT
95 Left GTTGAGGGTATTACTCCTGAGTGTG _15 Left CTCTCGTGTTTGTCCACTAAATGTG
96 Right CTGAGAACTGCAGCCTACAGAG _ 16 Right CTGAGAACTGCAGCCTACAGAGT
97 Left TGTTGAGGGTATTACTCCTGAGTGT _17 Left TTGACTCTGTCTCCTCTTGTCTTCT
98 Right CTGAGAACTGCAGCCTACAGAG _ 18 Right GAGGCTGTGAGCTGAGAACTG
99 Left TTGAGGGTATTACTCCTGAGTGTGT _19 Left CTCTCGTGTTTGTCCACTAAATGT
100 Right CTGAGAACTGCAGCCTACAGAG 10 Right AGAACTGCAGCCTACAGAGTCC
ALK Exon21 301-400 bases Seq.
Primer Sequence eq. Primer Sequence ID D
121 Left GAATCCTTCTTACCAGTTTTCAGGT 23 Left GTTGGAATCCTTCTTACCAGTTTTC
122 Right CTGAGAACTGCAGCCTACAGAGT 24 Right CTGAGAACTGCAGCCTACAGAGT

125 Left AATCCTTCTTACCAGTTTTCAGGTG 43 Left CAAAGCCATGTTGAGGGTATTACT
126 Right CTGAGAACTGCAGCCTACAGAGT 44 Right CTGAGAACTGCAGCCTACAGAGT
127 Left ATCCTTCTTACCAGTTTTCAGGTG 45 Left GAATCCTTCTTACCAGTTTTCAGGT
128 Right CTGAGAACTGCAGCCTACAGAGT 46 Right TGAGAACTGCAGCCTACAGAGT
129 Left GAATCCTTCTTACCAGTTTTCAGGT 47 Left GAATCCTTCTTACCAGTTTTCAGGT
130 Right AGAACTGCAGCCTACAGAGTCC 48 Right GAGAACTGCAGCCTACAGAGTCC
131 Left TTGGAATCCTTCTTACCAGTTTTC 49 Left GTTGGAATCCTTCTTACCAGTTTTC
132 Right CTGAGAACTGCAGCCTACAGAGT 50 Right CTGAGAACTGCAGCCTACAGAG
133 Left GTTGGAATCCTTCTTACCAGTTTTC 51 Left AATCCTTCTTACCAGTTTTCAGGTG
134 Right AGAACTGCAGCCTACAGAGTCC 52 Right AGAACTGCAGCCTACAGAGTCC
135 Left GAATCCTTCTTACCAGTTTTCAGG 53 Left ATCCTTCTTACCAGTTTTCAGGTG
136 Right CTGAGAACTGCAGCCTACAGAGT 54 Right AGAACTGCAGCCTACAGAGTCC
137 Left GGAATCCTTCTTACCAGTTTTCAG 55 Left GGTTGGAATCCTTCTTACCAGTTT
138 Right CTGAGAACTGCAGCCTACAGAGT 56 Right CTGAGAACTGCAGCCTACAGAGT
139 Left ATGTTGAGGGTATTACTCCTGAGTGT 57 Left TGGAATCCTTCTTACCAGTTTTCAG
140 Right CTGAGAACTGCAGCCTACAGAGT 58 Right CTGAGAACTGCAGCCTACAGAGT
141 Left GAATCCTTCTTACCAGTTTTCAGGT 59 Left TTGGAATCCTTCTTACCAGTTTTC
142 Right CTGAGAACTGCAGCCTACAGAG 60 Right AGAACTGCAGCCTACAGAGTCC
ALK Exon21-22 301-400 bases Seq. . 176 Right CCTTGGAGATATCGATCTGTTAGAA

Pnmer Sequence ID eq. .
161 Left TTGACTCTGTCTCCTCTTGTCTTCT ID Primer Sequence 162 Right TGGAGATATCGATCTGTTAGAAACC _ 77 Left GTTTGACTCTGTCTCCTCTTGTCTT
_ 78 Right TGGAGATATCGATCTGTTAGAAACC
163 Left TTTGACTCTGTCTCCTCTTGTCTTC
164 Right CCTTGGAGATATCGATCTGTTAGAA _ 79 Left TGACTCTGTCTCCTCTTGTCTTCTC
_ 80 Right TGGAGATATCGATCTGTTAGAAACC
165 Left ACTCTGTCTCCTCTTGTCTTCTC CT
166 Right CCTTGGAGATATCGATCTGTTAGAA _ 81 Left TGTTTGACTCTGTCTCCTCTTGTCT
_ 82 Right CCTTGGAGATATCGATCTGTTAGAA
167 Left GTTTGACTCTGTCTCCTCTTGTCTT
168 Right CCTTGGAGATATCGATCTGTTAGAA _ 83 Left CTGTTTGACTCTGTCTCCTCTTGTC
_ 84 Right CCTTGGAGATATCGATCTGTTAGAA
169 Left TTTGACTCTGTCTCCTCTTGTCTTC
170 Right TGGAGATATCGATCTGTTAGAAACC _ 85 Left CTCTGTCTCCTCTTGTCTTCTCCTT
_ 86 Right CCTTGGAGATATCGATCTGTTAGAA
171 Left ACTCTGTCTCCTCTTGTCTTCTC CT
172 Right TGGAGATATCGATCTGTTAGAAACC _ 87 Left TTTGACTCTGTCTCCTCTTGTCTTC
_ 88 Right TATCGATCTGTTAGAAACCTCTCCA
173 Left TTGACTCTGTCTCCTCTTGTCTTCT
174 Right TATCGATCTGTTAGAAACCTCTCCA _ 89 Left ACTCTGTCTCCTCTTGTCTTCTCCT
_ 90 Right TATCGATCTGTTAGAAACCTCTCCA
175 Left TGACTCTGTCTCCTCTTGTCTTCTC

191 Left TTGACTCTGTCTCCTCTTGTCTTCT 96 Right TGGAGATATCGATCTGTTAGAAACC
192 Right GTTAGAAACCTCTCCAGGTTCTTTG
_ 97 Left CTGTTTGACTCTGTCTCCTCTTGTC

Left GTTTGACTCTGTCTCCTCTTGTCTT _ 98 Right TGGAGATATCGATCTGTTAGAAACC
194 Right TATCGATCTGTTAGAAACCTCTCCA
_ 99 Left CTCTGTCTCCTCTTGTCTTCTCCTT
195 Left TGTTTGACTCTGTCTCCTCTTGTCT 200 Right TGGAGATATCGATCTGTTAGAAACC
ALK Exon21-22 401-500 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
201 Left TTGACTCTGTCTCCTCTTGTCTTCT 221 Left ACTCTGTCTCCTCTTGTCTTCTCCT
202 Right TAGAATGTTTGGGAGTCTCCTACTG 222 Right TAGAATGTTTGGGAGTCTCCTACTG
203 Left TTGACTCTGTCTCCTCTTGTCTTCT 223 Left TTGACTCTGTCTCCTCTTGTCTTCT
204 Right GTTGTTCCATTCTGGTAAGAAGTGT 224 Right TGTTCCATTCTGGTAAGAAGTGTCT
205 Left GTTGAGGGTATTACTCCTGAGTGTG 225 Left GTTTGACTCTGTCTCCTCTTGTCTT
206 Right CCTTGGAGATATCGATCTGTTAGAA 226 Right TAGAATGTTTGGGAGTCTCCTACTG
207 Left TGTTGAGGGTATTACTCCTGAGTGT 227 Left TTTGACTCTGTCTCCTCTTGTCTTC
208 Right CCTTGGAGATATCGATCTGTTAGAA 228 Right GTTGTTCCATTCTGGTAAGAAGTGT
209 Left TTGAGGGTATTACTCCTGAGTGTGT 229 Left ACTCTGTCTCCTCTTGTCTTCTCCT
210 Right CCTTGGAGATATCGATCTGTTAGAA 230 Right GTTGTTCCATTCTGGTAAGAAGTGT
211 Left GTTGAGGGTATTACTCCTGAGTGTG 231 Left GTTTGACTCTGTCTCCTCTTGTCTT
212 Right TGGAGATATCGATCTGTTAGAAACC 232 Right GTTGTTCCATTCTGGTAAGAAGTGT
213 Left TGTTGAGGGTATTACTCCTGAGTGT 233 Left TGACTCTGTCTCCTCTTGTCTTCTC
214 Right TGGAGATATCGATCTGTTAGAAACC 234 Right TAGAATGTTTGGGAGTCTCCTACTG
215 Left TTGAGGGTATTACTCCTGAGTGTGT 235 Left TGACTCTGTCTCCTCTTGTCTTCTC
216 Right TGGAGATATCGATCTGTTAGAAACC 236 Right GTTGTTCCATTCTGGTAAGAAGTGT
217 Left TTGACTCTGTCTCCTCTTGTCTTCT 237 Left GTTGAGGGTATTACTCCTGAGTGTG

Right GAAGTGTCTAGAATGTTTGGGAGTC 238 Right TATCGATCTGTTAGAAAC CTCTC CA
219 Left TTTGACTCTGTCTCCTCTTGTCTTC 239 Left TGTTGAGGGTATTACTCCTGAGTGT
220 Right TAGAATGTTTGGGAGTCTCCTACTG 240 Right TATCGATCTGTTAGAAAC CTCTC
CA
ALK Exon21-22 501-600 bases Seq. .
247 Left TGTTGAGGGTATTACTCCTGAGTGT
Pnmer Sequence ID
248 Right TAGAATGTTTGGGAGTCTCCTACTG
241 Left GAATCCTTCTTACCAGTTTTCAGGT seq.
242 Right CCTTGGAGATATCGATCTGTTAGAA Primer Sequence 249 Left TTGAGGGTATTACTCCTGAGTGTGT
243 Left GAATCCTTCTTACCAGTTTTCAGGT 250 Right TAGAATGTTTGGGAGTCTCCTACTG
244 Right TGGAGATATCGATCTGTTAGAAACC
251 Left GTTGAGGGTATTACTCCTGAGTGTG
245 Left GTTGAGGGTATTACTCCTGAGTGTG 252 Right GTTGTTCCATTCTGGTAAGAAGTGT
246 Right TAGAATGTTTGGGAGTCTCCTACTG
253 Left TGTTGAGGGTATTACTCCTGAGTGT

254 Right GTTGTTCCATTCTGGTAAGAAGTGT 267 Left GTTGAGGGTATTACTCCTGAGTGTG
268 Right TGTTCCATTCTGGTAAGAAGTGTCT
255 Left TTGAGGGTATTACTCCTGAGTGTGT
256 Right GTTGTTCCATTCTGGTAAGAAGTGT 269 Left TGTTGAGGGTATTACTCCTGAGTGT
270 Right TGTTCCATTCTGGTAAGAAGTGTCT
257 Left GTTGGAATCCTTCTTACCAGTTTTC
258 Right CCTTGGAGATATCGATCTGTTAGAA 271 Left TTGAGGGTATTACTCCTGAGTGTGT
272 Right TGTTCCATTCTGGTAAGAAGTGTCT
259 Left GTTGAGGGTATTACTCCTGAGTGTG
260 Right GAAGTGTCTAGAATGTTTGGGAGTC 273 Left AATCCTTCTTACCAGTTTTCAGGTG
274 Right CCTTGGAGATATCGATCTGTTAGAA
261 Left TTGAGGGTATTACTCCTGAGTGTGT
262 Right GAAGTGTCTAGAATGTTTGGGAGTC 275 Left GTTGGAATCCTTCTTACCAGTTTTC
276 Right TATCGATCTGTTAGAAACCTCTCCA
263 Left TGTTGAGGGTATTACTCCTGAGTGT
264 Right GAAGTGTCTAGAATGTTTGGGAGTC 277 Left ATCCTTCTTACCAGTTTTCAGGTG
278 Right CCTTGGAGATATCGATCTGTTAGAA
265 Left GAATCCTTCTTACCAGTTTTCAGGT
266 Right TATCGATCTGTTAGAAACCTCTCCA 279 Left GTTGAGGGTATTACTCCTGAGTGTG
280 Right TTGTTCCATTCTGGTAAGAAGTGTC
ALK Exon21-22 601-800 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
281 Left TTGACTCTGTCTCCTCTTGTCTTCT 301 Left ACTCTGTCTCCTCTTGTCTTCTCCT
282 Right AAAGTCTAGCATGCTCCATTTCTTA 302 Right AAAGTCTAGCATGCTCCATTTCTTA
283 Left GAATCCTTCTTACCAGTTTTCAGGT 303 Left TTGACTCTGTCTCCTCTTGTCTTCT
284 Right TAGAATGTTTGGGAGTCTCCTACTG 304 Right GGTCTTGGAGGGAGATTATATCTTG
285 Left GAATCCTTCTTACCAGTTTTCAGGT 305 Left GTTGGAATCCTTCTTACCAGTTTTC
286 Right AAAGTCTAGCATGCTCCATTTCTTA 306 Right TAGAATGTTTGGGAGTCTCCTACTG
287 Left GTTGAGGGTATTACTCCTGAGTGTG 307 Left GTTTGACTCTGTCTCCTCTTGTCTT
288 Right AAAGTCTAGCATGCTCCATTTCTTA 308 Right AAAGTCTAGCATGCTCCATTTCTTA
289 Left TGTTGAGGGTATTACTCCTGAGTGT 309 Left GAATCCTTCTTACCAGTTTTCAGGT
290 Right AAAGTCTAGCATGCTCCATTTCTTA 310 Right GAAGTGTCTAGAATGTTTGGGAGTC
291 Left TTGAGGGTATTACTCCTGAGTGTGT 311 Left CCTCTGTCACTCACTGGAAATACTC
292 Right AAAGTCTAGCATGCTCCATTTCTTA 312 Right TGGAGATATCGATCTGTTAGAAACC
293 Left CCTCTGTCACTCACTGGAAATACTC 313 Left TCCTCTGTCACTCACTGGAAATACT
294 Right CCTTGGAGATATCGATCTGTTAGAA 314 Right TGGAGATATCGATCTGTTAGAAACC
295 Left TCCTCTGTCACTCACTGGAAATACT 315 Left CTCTGTCACTCACTGGAAATACTCC
296 Right CCTTGGAGATATCGATCTGTTAGAA 316 Right TGGAGATATCGATCTGTTAGAAACC
297 Left CTCTGTCACTCACTGGAAATACTCC 317 Left GTTGGAATCCTTCTTACCAGTTTTC
298 Right CCTTGGAGATATCGATCTGTTAGAA 318 Right AAAGTCTAGCATGCTCCATTTCTTA
299 Left TTTGACTCTGTCTCCTCTTGTCTTC 319 Left TGACTCTGTCTCCTCTTGTCTTCTC
300 Right AAAGTCTAGCATGCTCCATTTCTTA 320 Right AAAGTCTAGCATGCTCCATTTCTTA

ALK Exon21-22 801-1000 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
321 Left CTCTCCTCAAAATTCATTCAGATGT 341 Left CTCTCCTCAAAATTCATTCAGATGT
322 Right CCTTGGAGATATCGATCTGTTAGAA 342 Right GAAGTGTCTAGAATGTTTGGGAGTC
323 Left ATGTTGGCTTACATTAACTCCCATA 343 Left CTCTCCTCAAAATTCATTCAGATGT
324 Right CCTTGGAGATATCGATCTGTTAGAA 344 Right TGTTCCATTCTGGTAAGAAGTGTCT
325 Left CTCTCCTCAAAATTCATTCAGATGT 345 Left CTCTCCTCAAAATTCATTCAGATGT
326 Right TAGAATGTTTGGGAGTCTCCTACTG 346 Right TATCGATCTGTTAGAAACCTCTCCA
327 Left ATGTTGGCTTACATTAACTCCCATA 347 Left ATGTTGGCTTACATTAACTCCCATA
328 Right TAGAATGTTTGGGAGTCTCCTACTG 348 Right TATCGATCTGTTAGAAACCTCTCCA
329 Left CTCTCCTCAAAATTCATTCAGATGT 349 Left AAAATTCATTCAGATGTGCTCTCTC
330 Right TGGAGATATCGATCTGTTAGAAACC 350 Right GAAGTGTCTAGAATGTTTGGGAGTC
331 Left CTCTCCTCAAAATTCATTCAGATGT 351 Left AAAATTCATTCAGATGTGCTCTCTC
332 Right GTTGTTCCATTCTGGTAAGAAGTGT 352 Right TGTTCCATTCTGGTAAGAAGTGTCT
333 Left ATGTTGGCTTACATTAACTCCCATA 353 Left TGGCTTACATTAACTCCCATAGTTT
334 Right TGGAGATATCGATCTGTTAGAAACC 354 Right CCTTGGAGATATCGATCTGTTAGAA
335 Left AAAATTCATTCAGATGTGCTCTCTC 355 Left TTGGCTTACATTAACTCCCATAGTT
336 Right TAGAATGTTTGGGAGTCTCCTACTG 356 Right CCTTGGAGATATCGATCTGTTAGAA
337 Left AAAATTCATTCAGATGTGCTCTCTC 357 Left TGTTGGCTTACATTAACTCCCATAG
338 Right TGGAGATATCGATCTGTTAGAAACC 358 Right CCTTGGAGATATCGATCTGTTAGAA
339 Left AAAATTCATTCAGATGTGCTCTCTC 359 Left AAAATTCATTCAGATGTGCTCTCTC
340 Right GTTGTTCCATTCTGGTAAGAAGTGT 360 Right TATCGATCTGTTAGAAACCTCTCCA
ALK Exon21-22 2kb S eq. . 372 Right CAAAGTTACATTTTCAGCAGCTACA

ID Pnmer Sequence 361 Left CTCTCCTCAAAATTCATTCAGATGT Seq. .
S
362 Right GCAGGAGAGTGTCTTTCTCAGATAC ID Primer equence 373 Left AAAATTCATTCAGATGTGCTCTCTC
363 Left ATGTTGGCTTACATTAACTCCCATA 374 Right GGAGAGTGTCTTTCTCAGATACTGG
364 Right GCAGGAGAGTGTCTTTCTCAGATAC
375 Left GAATCCTTCTTACCAGTTTTCAGGT
365 Left AAAATTCATTCAGATGTGCTCTCTC 376 Right CAAAGTTACATTTTCAGCAGCTACA
366 Right GCAGGAGAGTGTCTTTCTCAGATAC
377 Left CTCTCCTCAAAATTCATTCAGATGT
367 Left CTCTCCTCAAAATTCATTCAGATGT 378 Right GCAGCTACAATGTATAAAGGCATTC
368 Right GGAGAGTGTCTTTCTCAGATACTGG
379 Left GTTGAGGGTATTACTCCTGAGTGTG
369 Left ATGTTGGCTTACATTAACTCCCATA 380 Right CAAAGTTACATTTTCAGCAGCTACA
370 Right GGAGAGTGTCTTTCTCAGATACTGG
381 Left TGTTGAGGGTATTACTCCTGAGTGT
371 Left AAAATTCATTCAGATGTGCTCTCTC 382 Right CAAAGTTACATTTTCAGCAGCTACA

383 Left ATGTTGGCTTACATTAACTCCCATA 393 Left TCCTCTGTCACTCACTGGAAATACT
384 Right TTAACATGATCCCTTTAGGACACAC 394 Right GCAGGAGAGTGTCTTTCTCAGATAC
385 Left ATGTTGGCTTACATTAACTCCCATA 395 Left TGGCTTACATTAACTCCCATAGTTT
386 Right TGTTAACATGATCCCTTTAGGACAC 396 Right GCAGGAGAGTGTCTTTCTCAGATAC
387 Left ATGTTGGCTTACATTAACTCCCATA 397 Left TTGGCTTACATTAACTCCCATAGTT
388 Right GTTAACATGATCCCTTTAGGACACA 398 Right GCAGGAGAGTGTCTTTCTCAGATAC
389 Left AAAATTCATTCAGATGTGCTCTCTC 399 Left TGTTGGCTTACATTAACTCCCATAG
390 Right GCAGCTACAATGTATAAAGGCATTC 400 Right GCAGGAGAGTGTCTTTCTCAGATAC
391 Left CTCTGTCACTCACTGGAAATACTCC
392 Right GCAGGAGAGTGTCTTTCTCAGATAC
ALK Exon22 90-150 bases Seq.
Primer Sequence Primer Sequence ID ID
401 Left AGTTCTCAGCTCACAGCCTCCT 421 Left TAGGCTGCAGTTCTCAGCTCAC
402 Right AGGGTGTCTCTCTGTGGCTTTAC 422 Right AGGGTGTCTCTCTGTGGCTTTA
403 Left AGTTCTCAGCTCACAGCCTCCT 423 Left GCTGCAGTTCTCAGCTCACAG
404 Right GGGTGTCTCTCTGTGGCTTTAC 424 Right AGGGTGTCTCTCTGTGGCTTTAC
405 Left GTTCTCAGCTCACAGCCTCCT 425 Left TTCTCAGCTCACAGCCTCCT
406 Right AGGGTGTCTCTCTGTGGCTTTAC 426 Right AGGGTGTCTCTCTGTGGCTTTA
407 Left TAGGCTGCAGTTCTCAGCTCAC 427 Left GCTGCAGTTCTCAGCTCACAG
408 Right AGGGTGTCTCTCTGTGGCTTTAC 428 Right GGGTGTCTCTCTGTGGCTTTAC
409 Left GTTCTCAGCTCACAGCCTCCT 429 Left AGTTCTCAGCTCACAGCCTCCT
410 Right GGGTGTCTCTCTGTGGCTTTAC 430 Right GAGGGTGTCTCTCTGTGGCTTTAC
411 Left AGTTCTCAGCTCACAGCCTCCT 431 Left AGTTCTCAGCTCACAGCCTCCTC
412 Right AGGGTGTCTCTCTGTGGCTTTA 432 Right AGGGTGTCTCTCTGTGGCTTTAC
413 Left TAGGCTGCAGTTCTCAGCTCAC 433 Left GTTCTCAGCTCACAGCCTCCT
414 Right GGGTGTCTCTCTGTGGCTTTAC 434 Right GAGGGTGTCTCTCTGTGGCTTTAC
415 Left TTCTCAGCTCACAGCCTCCT 435 Left TAGGCTGCAGTTCTCAGCTCAC
416 Right AGGGTGTCTCTCTGTGGCTTTAC 436 Right GAGGGTGTCTCTCTGTGGCTTTAC
417 Left GTTCTCAGCTCACAGCCTCCT 437 Left AGTTCTCAGCTCACAGCCTCCTC
418 Right AGGGTGTCTCTCTGTGGCTTTA 438 Right GGGTGTCTCTCTGTGGCTTTAC
419 Left TTCTCAGCTCACAGCCTCCT 439 Left TCTCAGCTCACAGCCTCCTC
420 Right GGGTGTCTCTCTGTGGCTTTAC 440 Right AGGGTGTCTCTCTGTGGCTTTAC
ALK Exon22 151-200 bases S eq. . 442 Right TATCGATCTGTTAGAAAC CTCTC
CA
ID Pnmer Sequence 441 Left AGTTCTCAGCTCACAGCCTCCT 443 Left GTTCTCAGCTCACAGCCTCCT

444 Right TATCGATCTGTTAGAAACCTCTCCA
Seq. 463 Left GTTCTCAGCTCACAGCCTCCT
Primer Sequence ID ht GATATCGATCTGTTAGAAACCTCTC
6 Rig 445 Left TTCTCAGCTCACAGCCTCCT C
446 Right TATCGATCTGTTAGAAACCTCTCCA
465 Left GTTCTCAGCTCACAGCCTCCT
447 Left AGTTCTCAGCTCACAGCCTCCT 466 Right TTAGAAACCTCTCCAGGTTCTTTG
448 Right GTTAGAAACCTCTCCAGGTTCTTTG
467 Left AGTTCTCAGCTCACAGCCTCCT
449 Left GTTCTCAGCTCACAGCCTCCT 468 Right ATCGATCTGTTAGAAACCTCTCCAG
450 Right GTTAGAAACCTCTCCAGGTTCTTTG
469 Left TAGGCTGCAGTTCTCAGCTCAC
451 Left TCTCAGCTCACAGCCTCCTC 470 Right TTAGAAACCTCTCCAGGTTCTTTG

452 Right TGGAGATATCGATCTGTTAGAAACC
471 Left AGCTCACAGCCTCCTCCTC
453 Left TAGGCTGCAGTTCTCAGCTCAC 472 Right CCTTGGAGATATCGATCTGTTAGAA
454 Right GTTAGAAACCTCTCCAGGTTCTTTG
473 Left GCTGCAGTTCTCAGCTCACAG
455 Left AGTTCTCAGCTCACAGCCTCCT 474 Right GTTAGAAACCTCTCCAGGTTCTTTG
GATATCGATCTGTTAGAAACCTCTC
_____________________________________________________________ 456 Right C 475 Left TCTCAGCTCACAGCCTCCTC
¨476 Right TATCGATCTGTTAGAAACCTCTCCA
457 Left TTCTCAGCTCACAGCCTCCT
¨
458 Right GTTAGAAACCTCTCCAGGTTCTTTG 477 Left TTCTCAGCTCACAGCCTCCT
__________________________________________ 4 78 Right GATATCGATCTGTTAGAAACCTCTC
459 Left AGTTCTCAGCTCACAGCCTCCT C
460 Right TTAGAAACCTCTCCAGGTTCTTTG
479 Left GTTCTCAGCTCACAGCCTCCTC
461 Left AGTTCTCAGCTCACAGCCTCCTC 480 Right TATCGATCTGTTAGAAACCTCTCCA
462 Right TATCGATCTGTTAGAAACCTCTCCA
ALK Exon22 201-300 bases Seq.
Primer Sequence ID 495 Left GGACTCTGTAGGCTGCAGTTCTC
481 Left GGACTCTGTAGGCTGCAGTTCTC 4 96 Right TGGTAAGAAGTGTCTAGAATGTTTG
482 Right CCTTGGAGATATCGATCTGTTAGAA G
__________________________________________ Seq.
483 Left GGACTCTGTAGGCTGCAGTTCTC ID Primer Sequence 484 Right TAGAATGTTTGGGAGTCTCCTACTG 497 Left GGACTCTGTAGGCTGCAGTTCTC
¨498 Right AGAATGTTTGGGAGTCTCCTACTG
485 Left GGACTCTGTAGGCTGCAGTTCTC ________ ¨
486 Right TGGAGATATCGATCTGTTAGAAACC499 Left GGACTCTGTAGGCTGCAGTTCTC
500 Right TCTAGAATGTTTGGGAGTCTCCTACT
487 Left GGACTCTGTAGGCTGCAGTTCTC
488 Right GAAGTGTCTAGAATGTTTGGGAGTC 501 Left GGACTCTGTAGGCTGCAGTTCTC
__________________________________________ 502 R ht GATATCGATCTGTTAGAAACCTCTC
489 Left GGACTCTGTAGGCTGCAGTTCTC C
490 Right TATCGATCTGTTAGAAACCTCTCCA _______ __________________________________________ 503 Left GGACTCTGTAGGCTGCAGTTCTC
491 Left GGACTCTGTAGGCTGCAGTTCTC 504 Right TTAGAAACCTCTCCAGGTTCTTTG
492 Right GTTAGAAACCTCTCCAGGTTCTTTG _______ __________________________________________ 505 Left GGACTCTGTAGGCTGCAGTTCTC
493 Left GGACTCTGTAGGCTGCAGTTCTC 506 Right AGAAGTGTCTAGAATGTTTGGGAGT
4 94 Right AAGAAGTGTCTAGAATGTTTGGGAG
__________________________________________________ T 507 Left GGACTCTGTAGGCTGCAGTTCTC

508 Right AAGTGTCTAGAATGTTTGGGAGTCT
515 Left GGACTCTGTAGGCTGCAGTTCTC
509 Left AGTTCTCAGCTCACAGCCTCCT 516 Right ATCGATCTGTTAGAAACCTCTCCAG
510 Right CCTTGGAGATATCGATCTGTTAGAA
517 Left GTTCTCAGCTCACAGCCTCCT
511 Left AGTTCTCAGCTCACAGCCTCCT 518 Right CCTTGGAGATATCGATCTGTTAGAA
512 Right TAGAATGTTTGGGAGTCTCCTACTG
519 Left AGTTCTCAGCTCACAGCCTCCT
513 Left AGTTCTCAGCTCACAGCCTCCT 520 Right GTTGTTCCATTCTGGTAAGAAGTGT
514 Right TGGAGATATCGATCTGTTAGAAACC
ALK Exon22 301-400 bases Seq.
Primer Sequence Primer Sequence ID ID
521 Left GGACTCTGTAGGCTGCAGTTCTC 541 Left GGACTCTGTAGGCTGCAGTTCTC
522 Right GTTGTTCCATTCTGGTAAGAAGTGT 542 Right TTGTTCCATTCTGGTAAGAAGTGT
523 Left GGACTCTGTAGGCTGCAGTTCTC 543 Left GGACTCTGTAGGCTGCAGTTCTC
524 Right TGTTCCATTCTGGTAAGAAGTGTCT 544 Right GTTCCATTCTGGTAAGAAGTGTCTA
525 Left GGACTCTGTAGGCTGCAGTTCTC 545 Left GGACTCTGTAGGCTGCAGTTCTC
526 Right TTGTTCCATTCTGGTAAGAAGTGTC 546 Right GATACTGGTTGCAGACAGTGACATC
527 Left GGACTCTGTAGGCTGCAGTTCTC 547 Left CGGACTCTGTAGGCTGCAGTT
528 Right GGTTGTTCCATTCTGGTAAGAAGT 548 Right GTTGTTCCATTCTGGTAAGAAGTGT
529 Left GGACTCTGTAGGCTGCAGTTCTC 549 Left GGACTCTGTAGGCTGCAGTTCTC
530 Right GGATTATTAGGCCACACAGACTTT 550 Right TAGGCCACACAGACTTTGTTTCT
531 Left GGACTCTGTAGGCTGCAGTTCTC 551 Left GGACTCTGTAGGCTGCAGTTCTC
TGTTCCATTCTGGTAAGAAGTGTCT 552 Right TACTGGTTGCAGACAGTGACATC
532 Right A
553 Left GTAGGCTGCAGTTCTCAGCTCACAG
533 Left GGACTCTGTAGGCTGCAGTTCTC 554 Right GTTGTTCCATTCTGGTAAGAAGTGT
534 Right TGTTCCATTCTGGTAAGAAGTGTC _ __________________________________________________ 555 Left AGTTCTCAGCTCACAGCCTCCT
535 Left GGACTCTGTAGGCTGCAGTTCTC 556 Right GGATTATTAGGCCACACAGACTTT
536 Right ATACTGGTTGCAGACAGTGACATC
557 Left GGACTCTGTAGGCTGCAGTTCTC
537 Left GGACTCTGTAGGCTGCAGTTCTC 558 Right TTAGGCCACACAGACTTTGTTTCT
538 Right GATACTGGTTGCAGACAGTGACAT
559 Left GGACTCTGTAGGCTGCAGTTCTC
539 Left GGACTCTGTAGGCTGCAGTTCTC 560 Right ACAGTGACATCGGTGGGATTATTAG
540 Right ATTAGGCCACACAGACTTTGTTTCT
ALK Exon23 151-200 bases S eq. 566 Right AGCAAAGACTGGTTCTCACTCAC

ID Primer Sequence 561 Left TTAATTTTGGTTACATCCCTCTCTG _567 Left AGACTCAGCTCAGTTAATTTTGGTT
562 Right AGCAAAGACTGGTTCTCACTCAC 568 Right AGCAAAGACTGGTTCTCACTCAC
___________________________________________ Seq.
563 Left CAGACTCAGCTCAGTTAATTTTGGT ID Primer Sequence 564 Right AGCAAAGACTGGTTCTCACTCAC 569 Left CTCAGCTCAGTTAATTTTGGTTACA
570 Right AGCAAAGACTGGTTCTCACTCAC
565 Left AGCTCAGTTAATTTTGGTTACATCC

571 Left TCAGCTCAGTTAATTTTGGTTACATC 586 Right AGCAAAGACTGGTTCTCACTCAC
572 Right AGCAAAGACTGGTTCTCACTCAC
587 Left TCAGCTCAGTTAATTTTGGTTACAT
573 Left CAGTTAATTTTGGTTACATCCCTCT 588 Right AGCAAAGACTGGTTCTCACTCAC
574 Right AGCAAAGACTGGTTCTCACTCAC
589 Left TTAATTTTGGTTACATCCCTCTCTG
575 Left ACTCAGCTCAGTTAATTTTGGTTACA 590 Right AACTGCAGCAAAGACTGGTTCT
576 Right AGCAAAGACTGGTTCTCACTCAC
591 Left TTAATTTTGGTTACATCCCTCTCTG
577 Left CAGTTAATTTTGGTTACATCCCTCTC 592 Right ACAACAACTGCAGCAAAGACTG
578 Right AGCAAAGACTGGTTCTCACTCAC
593 Left TTAATTTTGGTTACATCCCTCTCTG
579 Left TCAGTTAATTTTGGTTACATCCCTCT 594 Right CACAACAACTGCAGCAAAGACT
580 Right AGCAAAGACTGGTTCTCACTCAC
595 Left CTCAGCTCAGTTAATTTTGGTTACAT
581 Left GTTAATTTTGGTTACATCCCTCTCTG 596 Right AGCAAAGACTGGTTCTCACTCAC
582 Right AGCAAAGACTGGTTCTCACTCAC
597 Left TTAATTTTGGTTACATCCCTCTCTG
583 Left CAGACTCAGCTCAGTTAATTTTGG 598 Right CAGCAAAGACTGGTTCTCACTCAC
584 Right AGCAAAGACTGGTTCTCACTCAC
599 Left AGTTAATTTTGGTTACATCCCTCTC
585 Left CAGCTCAGTTAATTTTGGTTACATC ,500 Right AGCAAAGACTGGTTCTCACTCAC
ALK Exon23 201-300 bases Seq. . L620 Right GTCCAAGCCTAAAGTTGACACC
ID
Pnmer Sequence 601 Left TGTAGCTGCTGAAAATGTAACTTTG
602 Right AGCAAAGACTGGTTCTCACTCAC
__________________________________________ Seq.
603 Left TTAATTTTGGTTACATCCCTCTCTG ID Primer Sequence 604 Right CTGTCCAAGCCTAAAGTTGACAC ¨621 Left CAGTTAATTTTGGTTACATCCCTCT
_________________________________________ _622 Right CTGTCCAAGCCTAAAGTTGACAC
605 Left TATCCTGTTCCTCCCAGTTTAAGAT ____ 606 Right AGCAAAGACTGGTTCTCACTCAC 623 Left TTAATTTTGGTTACATCCCTCTCTG
_________________________________________ _624 Right CCTGTCCAAGCCTAAAGTTGAC
607 Left ATGCCTTTATACATTGTAGCTGCTG ________ 608 Right AGCAAAGACTGGTTCTCACTCAC _625 Left TATCCTGTTCCTCCCAGTTTAAGA
_________________________________________ _626 Right AGCAAAGACTGGTTCTCACTCAC
609 Left GCCTTTATACATTGTAGCTGCTGAA ________ 610 Right AGCAAAGACTGGTTCTCACTCAC _627 Left CTGTTCCTCCCAGTTTAAGATTTG
__________________________________________ 628 Right AGCAAAGACTGGTTCTCACTCAC
611 Left GTATCCTGTTCCTCCCAGTTTAAGA ________ 612 Right AGCAAAGACTGGTTCTCACTCAC _629 Left CAGAATGCCTTTATACATTGTAGCTG
__________________________________________ 630 Right AGCAAAGACTGGTTCTCACTCAC
613 Left GCCTTTATACATTGTAGCTGCTGA _______ 614 Right AGCAAAGACTGGTTCTCACTCAC 631 Left CCCATGTTTACAGAATGCCTTTAT
__________________________________________ 632 Right AGCAAAGACTGGTTCTCACTCAC
615 Left AGTTTAAGATTTGCCCAGACTCAG _______ 616 Right AGCAAAGACTGGTTCTCACTCAC 633 Left GCTGCTGAAAATGTAACTTTGTATC
_________________________________________ _634 Right AGCAAAGACTGGTTCTCACTCAC
617 Left CCCAGACTCAGCTCAGTTAATTTT ________ 618 Right AGCAAAGACTGGTTCTCACTCAC 635 Left GTATCCTGTTCCTCCCAGTTTAAG
__________________________________________ 636 Right AGCAAAGACTGGTTCTCACTCAC
619 Left TTAATTTTGGTTACATCCCTCTCTG

637 Left TGTAGCTGCTGAAAATGTAACTTTG 639 Left ATCCTGTTCCTCCCAGTTTAAGAT
638 Right AACTGCAGCAAAGACTGGTTCT ,540 Right AGCAAAGACTGGTTCTCACTCAC

ALK Exon23 301-400 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
641 Left TTAATTTTGGTTACATCCCTCTCTG 561 Left CAGACTCAGCTCAGTTAATTTTGGT
642 Right TCAGCCATCATCTACCTCTATCTTC 562 Right CAGCCATCATCTACCTCTATCTTCT
643 Left CAGACTCAGCTCAGTTAATTTTGGT 563 Left CAGACTCAGCTCAGTTAATTTTGGT
644 Right TCAGCCATCATCTACCTCTATCTTC 564 Right CTCAGCCATCATCTACCTCTATCTT
645 Left TTAATTTTGGTTACATCCCTCTCTG 565 Left CAGACTCAGCTCAGTTAATTTTGGT
646 Right CTATCTTCTGTCCATTCTCTTCCAG 566 Right AGCCATCATCTACCTCTATCTTCTG
647 Left CAGACTCAGCTCAGTTAATTTTGGT 567 Left TTAATTTTGGTTACATCCCTCTCTG
648 Right CTATCTTCTGTCCATTCTCTTCCAG 568 Right GCCATCATCTACCTCTATCTTCTGT
649 Left TATCCTGTTCCTCCCAGTTTAAGAT 569 Left CAGACTCAGCTCAGTTAATTTTGGT
650 Right CTATCTTCTGTCCATTCTCTTCCAG 570 Right GCCATCATCTACCTCTATCTTCTGT
651 Left TTAATTTTGGTTACATCCCTCTCTG 571 Left AGCTCAGTTAATTTTGGTTACATCC
652 Right TCTATCTTCTGTCCATTCTCTTCCA 572 Right TCAGCCATCATCTACCTCTATCTTC
653 Left CAGACTCAGCTCAGTTAATTTTGGT 573 Left AGCTCAGTTAATTTTGGTTACATCC
654 Right TCTATCTTCTGTCCATTCTCTTCCA 574 Right CTATCTTCTGTCCATTCTCTTCCAG
655 Left TTAATTTTGGTTACATCCCTCTCTG 575 Left AGACTCAGCTCAGTTAATTTTGGTT
656 Right CAGCCATCATCTACCTCTATCTTCT 576 Right TCAGCCATCATCTACCTCTATCTTC
657 Left TTAATTTTGGTTACATCCCTCTCTG 577 Left CTCAGCTCAGTTAATTTTGGTTACA
658 Right CTCAGCCATCATCTACCTCTATCTT 578 Right TCAGCCATCATCTACCTCTATCTTC
659 Left TTAATTTTGGTTACATCCCTCTCTG 579 Left TATCCTGTTCCTCCCAGTTTAAGAT
660 Right AGCCATCATCTACCTCTATCTTCTG 580 Right TCTATCTTCTGTCCATTCTCTTCCA
ALK Exon23 401-600 bases S eq. . 590 Right CTTCTGCAATGATTGTAAGTTTCCT
ID Pnmer Sequence 681 Left TGTAGCTGCTGAAAATGTAACTTTG 591 Left TGTAGCTGCTGAAAATGTAACTTTG
682 Right TCAGCCATCATCTACCTCTATCTTC 592 Right CTATCTTCTGTCCATTCTCTTCCAG
__________________________________________ Seq. .
683 Left TTAATTTTGGTTACATCCCTCTCTG ID Primer Sequence 684 Right ACCTTCTGCAATGATTGTAAGTTTC 593 Left TATCCTGTTCCTCCCAGTTTAAGAT
594 Right ACCTTCTGCAATGATTGTAAGTTTC
685 Left CAGACTCAGCTCAGTTAATTTTGGT
686 Right ACCTTCTGCAATGATTGTAAGTTTC 595 Left TATCCTGTTCCTCCCAGTTTAAGAT
596 Right CTTCTGCAATGATTGTAAGTTTCCT
687 Left TTAATTTTGGTTACATCCCTCTCTG
688 Right CTTCTGCAATGATTGTAAGTTTCCT 597 Left TGTAGCTGCTGAAAATGTAACTTTG
598 Right TCTATCTTCTGTCCATTCTCTTCCA
689 Left CAGACTCAGCTCAGTTAATTTTGGT

699 Left TGTAGCTGCTGAAAATGTAACTTTG 710 Right ACCTTCTGCAATGATTGTAAGTTTC
700 Right CAGCCATCATCTACCTCTATCTTCT
711 Left CAGACTCAGCTCAGTTAATTTTGGT
701 Left TGTAGCTGCTGAAAATGTAACTTTG 712 Right CACCTTCTGCAATGATTGTAAGTTT
702 Right AGCCATCATCTACCTCTATCTTCTG
713 Left AGCTCAGTTAATTTTGGTTACATCC
703 Left TGTAGCTGCTGAAAATGTAACTTTG 714 Right CTTCTGCAATGATTGTAAGTTTCCT
704 Right CTCAGCCATCATCTACCTCTATCTT
715 Left ATGCCTTTATACATTGTAGCTGCTG
705 Left TGTAGCTGCTGAAAATGTAACTTTG 716 Right TCAGCCATCATCTACCTCTATCTTC
706 Right GCCATCATCTACCTCTATCTTCTGT
717 Left TGTAGCTGCTGAAAATGTAACTTTG
707 Left TTAATTTTGGTTACATCCCTCTCTG 718 Right GAGCCACTTAAATCTCTTTTCTTTG
708 Right CACCTTCTGCAATGATTGTAAGTTT
719 Left TGTAGCTGCTGAAAATGTAACTTTG
709 Left AGCTCAGTTAATTTTGGTTACATCC 720 Right TGAGCCACTTAAATCTCTTTTCTTT
ALK Exon23 601-800 bases Seq.
Primer Sequence Primer Sequence ID ID
721 Left TGTAGCTGCTGAAAATGTAACTTTG 741 Left TTAATTTTGGTTACATCCCTCTCTG
722 Right ACCTTCTGCAATGATTGTAAGTTTC 742 Right TAGAATGTGGATATGGTTTGGATTT
723 Left TGTAGCTGCTGAAAATGTAACTTTG 743 Left CAGACTCAGCTCAGTTAATTTTGGT
724 Right GTTGAATTGTAATCCCTAGTGTTGG 744 Right TAGAATGTGGATATGGTTTGGATTT
725 Left TGTAGCTGCTGAAAATGTAACTTTG 745 Left CCAGTATCTGAGAAAGACACTCTCC
726 Right CTTCTGCAATGATTGTAAGTTTCCT 746 Right CTATCTTCTGTCCATTCTCTTCCAG
727 Left TTAATTTTGGTTACATCCCTCTCTG 747 Left TTAATTTTGGTTACATCCCTCTCTG
728 Right GCTATAGAATGTGGATATGGTTTGG 748 Right TCATGAGACCTGGTTGTTTAAAAGT
729 Left TTAATTTTGGTTACATCCCTCTCTG 749 Left TTAATTTTGGTTACATCCCTCTCTG
730 Right GGCTATAGAATGTGGATATGGTTTG 750 Right CAGGCTATAGAATGTGGATATGGTT
731 Left CAGACTCAGCTCAGTTAATTTTGGT 751 Left TGTAGCTGCTGAAAATGTAACTTTG
732 Right GCTATAGAATGTGGATATGGTTTGG 752 Right CTCATGAGACCTGGTTGTTTAAAAG
733 Left CAGACTCAGCTCAGTTAATTTTGGT 753 Left CAGACTCAGCTCAGTTAATTTTGGT
734 Right GGCTATAGAATGTGGATATGGTTTG 754 Right TCATGAGACCTGGTTGTTTAAAAGT
735 Left TTAATTTTGGTTACATCCCTCTCTG 755 Left CAGACTCAGCTCAGTTAATTTTGGT
736 Right GTTGAATTGTAATCCCTAGTGTTGG 756 Right CAGGCTATAGAATGTGGATATGGTT
737 Left CAGACTCAGCTCAGTTAATTTTGGT 757 Left TTAATTTTGGTTACATCCCTCTCTG
738 Right GTTGAATTGTAATCCCTAGTGTTGG 758 Right ATCCAGGCTATAGAATGTGGATATG
739 Left CCAGTATCTGAGAAAGACACTCTCC 759 Left CAGACTCAGCTCAGTTAATTTTGGT
740 Right TCAGCCATCATCTACCTCTATCTTC 760 Right ATCCAGGCTATAGAATGTGGATATG
ALK Exon23 801-1000 bases S eq.761 Left TGTAGCTGCTGAAAATGTAACTTTG
Primer Sequence ID 762 Right GCTATAGAATGTGGATATGGTTTGG

782 Right TCAGCCATCATCTACCTCTATCTTC
763 Left TGTAGCTGCTGAAAATGTAACTTTG
764 Right GGCTATAGAATGTGGATATGGTTTG 783 Left TTAATTTTGGTTACATCCCTCTCTG
S eq. . 784 Right TATTATCCCTACTTGAGACGTGAGG
ID Pnmer Sequence ¨
765 Left TGTAGCTGCTGAAAATGTAACTTTG 785 Left TTAATTTTGGTTACATCCCTCTCTG
766 Right GTCATGAAAGTTCTCCTCTGTGTTT 786 Right CAGGTCAGTTGCTTGAGTAGTTACA
767 Left TGTAGCTGCTGAAAATGTAACTTTG 787 Left TTAATTTTGGTTACATCCCTCTCTG
768 Right ATGAAAGTTCTCCTCTGTGTTTGTC _788 Right GTCATGAAAGTTCTCCTCTGTGTTT
769 Left CCAGTATCTGAGAAAGACACTCTCC 789 Left TTAATTTTGGTTACATCCCTCTCTG
770 Right ACCTTCTGCAATGATTGTAAGTTTC 790 Right ATGAAAGTTCTCCTCTGTGTTTGTC
771 Left TGTAGCTGCTGAAAATGTAACTTTG 791 Left CAGACTCAGCTCAGTTAATTTTGGT
772 Right TAGAATGTGGATATGGTTTGGATTT 792 Right TATTATCCCTACTTGAGACGTGAGG
773 Left TGTAGCTGCTGAAAATGTAACTTTG 793 Left CAGACTCAGCTCAGTTAATTTTGGT
774 Right CTCTAGTTTGGTTTTCCAGAGTCAG 794 Right CAGGTCAGTTGCTTGAGTAGTTACA
775 Left TGTAGCTGCTGAAAATGTAACTTTG 795 Left CAGACTCAGCTCAGTTAATTTTGGT
776 Right CCTCTGTGTTTGTCTCTAGTTTGGT 796 Right GTCATGAAAGTTCTCCTCTGTGTTT
777 Left CCAGTATCTGAGAAAGACACTCTCC 797 Left CAGACTCAGCTCAGTTAATTTTGGT
778 Right CTTCTGCAATGATTGTAAGTTTCCT 798 Right ATGAAAGTTCTCCTCTGTGTTTGTC
779 Left TGTAGCTGCTGAAAATGTAACTTTG 799 Left TTAATTTTGGTTACATCCCTCTCTG
780 Right GAAAGTTCTCCTCTGTGTTTGTCTC 800 Right CTCTAGTTTGGTTTTCCAGAGTCAG
781 Left CATGTTAACAAGAAAACCCAAGTCT
ALK Exon23 2kb S eq. . 314 Right CTTCTGCAATGATTGTAAGTTTCCT
ID Pnmer Sequence 801 Left TTCTAACAGATCGATATCTCCAAGG 315 Left GAGCATGCTAGACTTTGACAGTACA
802 Right GCTATAGAATGTGGATATGGTTTGG 316 Right GTTGAATTGTAATCCCTAGTGTTGG
803 Left GAGCATGCTAGACTTTGACAGTACA 217 Left ACAGGAGGACACACAAAATAACATT
804 Right GCTATAGAATGTGGATATGGTTTGG 318 Right ATAGTACAGTGGTTCGTTGAGGAAG
__________________________________________ Seq.
Primer Sequence 805 Left GAGCATGCTAGACTTTGACAGTACA ID
806 Right GGCTATAGAATGTGGATATGGTTTG 319 Left GAGCATGCTAGACTTTGACAGTACA
320 Right TATTATCCCTACTTGAGACGTGAGG
807 Left TTCTAACAGATCGATATCTCCAAGG
808 Right ACCTTCTGCAATGATTGTAAGTTTC 321 Left GAGCATGCTAGACTTTGACAGTACA
322 Right CAGGTCAGTTGCTTGAGTAGTTACA
809 Left GAGCATGCTAGACTTTGACAGTACA
810 Right CCAAGCTCTGTTAACCATAAGATGT 323 Left GAGCATGCTAGACTTTGACAGTACA
324 Right GTCATGAAAGTTCTCCTCTGTGTTT
811 Left TTCTAACAGATCGATATCTCCAAGG
812 Right GTTGAATTGTAATCCCTAGTGTTGG 325 Left AGATATTTGACCTCAAGATCAGGTG
326 Right AGGTTAAAGGTTTAAGACTGCCCTA
813 Left TTCTAACAGATCGATATCTCCAAGG

827 Left CAGTAGGAGACTCCCAAACATTCTA 334 Right TAGAATGTGGATATGGTTTGGATTT
828 Right GTTGAATTGTAATCCCTAGTGTTGG
335 Left CATGTTAACAAGAAAACCCAAGTCT
829 Left ACAGGAGGACACACAAAATAACATT 336 Right ATAGTACAGTGGTTCGTTGAGGAAG
830 Right AGGTTAAAGGTTTAAGACTGCCCTA
337 Left AGATATTTGACCTCAAGATCAGGTG
831 Left AGATATTTGACCTCAAGATCAGGTG 338 Right CACCTTTGAGATGTTCTAGTCCAAT
832 Right CCAAGCTCTGTTAACCATAAGATGT
339 Left ACAGGAGGACACACAAAATAACATT
833 Left TTCTAACAGATCGATATCTCCAAGG 340 Right CACCTTTGAGATGTTCTAGTCCAAT
ALK Exon21-23 2kb Seq. Seq.
Primer Sequence Primer Sequence ID ID
841 Left TTGACTCTGTCTCCTCTTGTCTTCT 861 Left TTGACTCTGTCTCCTCTTGTCTTCT
842 Right TCAGCCATCATCTACCTCTATCTTC 862 Right CAGCCATCATCTACCTCTATCTTCT
843 Left TTGACTCTGTCTCCTCTTGTCTTCT 863 Left TTGACTCTGTCTCCTCTTGTCTTCT
844 Right CTATCTTCTGTCCATTCTCTTCCAG 864 Right CTCAGCCATCATCTACCTCTATCTT
845 Left TTTGACTCTGTCTCCTCTTGTCTTC 865 Left TTGACTCTGTCTCCTCTTGTCTTCT
846 Right TCAGCCATCATCTACCTCTATCTTC 866 Right AGCCATCATCTACCTCTATCTTCTG
847 Left ACTCTGTCTCCTCTTGTCTTCTCCT 867 Left TGACTCTGTCTCCTCTTGTCTTCTC
848 Right TCAGCCATCATCTACCTCTATCTTC 868 Right CTATCTTCTGTCCATTCTCTTCCAG
849 Left GTTTGACTCTGTCTCCTCTTGTCTT 869 Left TTGACTCTGTCTCCTCTTGTCTTCT
850 Right TCAGCCATCATCTACCTCTATCTTC 870 Right GCCATCATCTACCTCTATCTTCTGT
851 Left TTTGACTCTGTCTCCTCTTGTCTTC 871 Left TGTTTGACTCTGTCTCCTCTTGTCT
852 Right CTATCTTCTGTCCATTCTCTTCCAG 872 Right TCAGCCATCATCTACCTCTATCTTC
853 Left ACTCTGTCTCCTCTTGTCTTCTCCT 873 Left CTGTTTGACTCTGTCTCCTCTTGTC
854 Right CTATCTTCTGTCCATTCTCTTCCAG 874 Right TCAGCCATCATCTACCTCTATCTTC
855 Left GTTTGACTCTGTCTCCTCTTGTCTT 875 Left CTCTGTCTCCTCTTGTCTTCTCCTT
856 Right CTATCTTCTGTCCATTCTCTTCCAG 876 Right TCAGCCATCATCTACCTCTATCTTC
857 Left TTGACTCTGTCTCCTCTTGTCTTCT 877 Left TTTGACTCTGTCTCCTCTTGTCTTC
858 Right TCTATCTTCTGTCCATTCTCTTCCA 878 Right TCTATCTTCTGTCCATTCTCTTCCA
859 Left TGACTCTGTCTCCTCTTGTCTTCTC 879 Left TGTTTGACTCTGTCTCCTCTTGTCT
860 Right TCAGCCATCATCTACCTCTATCTTC 880 Right CTATCTTCTGTCCATTCTCTTCCAG
ALK Exon21-23 5kb Seq. . 885 Left CAGTGTAGGGGCTGAATGTTATC
Primer Sequence ID 886 Right ATACAACTTTCTCTCCTTAAGCCTCA
881 Left CAGTGTAGGGGCTGAATGTTATC
882 Right TACAACTTTCTCTCCTTAAGCCTCA 887 Left GCAGTGTAGGGGCTGAATGTTAT
888 Right TACAACTTTCTCTCCTTAAGCCTCA
883 Left CAGTGTAGGGGCTGAATGTTATC
884 Right ACAACTTTCTCTCCTTAAGCCTCA _889 Left CAGACTCCTCTAGCCACAAAAGG
890 Right TACAACTTTCTCTCCTTAAGCCTCA

Seq. . 905 Left ATGTTGGCTTACATTAACTCCCATA

Primer Sequence ID 906 Right TTTGCAAAGTCCCTCTCCTTT
891 Left GCAGACTCCTCTAGCCACAAAAG ________ 892 Right TACAACTTTCTCTCCTTAAGCCTCA 907 Left GTTATCACAGCACCGCAGACT
_908 Right TACAACTTTCTCTCCTTAAGCCTCA
893 Left GCAGACTCCTCTAGCCACAAAA
894 Right TACAACTTTCTCTCCTTAAGCCTCA 909 Left GCAGACTCCTCTAGCCACAAA
_________________________________________ _910 Right TACAACTTTCTCTCCTTAAGCCTCA

895 Left AGACTCCTCTAGCCACAAAAGG
896 Right TACAACTTTCTCTCCTTAAGCCTCA _911 Left GCAGTGTAGGGGCTGAATGTTA
912 Right TACAACTTTCTCTCCTTAAGCCTCA
897 Left CAGTGTAGGGGCTGAATGTTATC
898 Right GCATGCATACAACTTTCTCTCCTT 913 Left TAGGGGCTGAATGTTATCACAGC
914 Right TACAACTTTCTCTCCTTAAGCCTCA
899 Left GCAGTGTAGGGGCTGAATGTTATC
900 Right TACAACTTTCTCTCCTTAAGCCTCA 915 Left CAGACTCCTCTAGCCACAAAAGG
916 Right GCATGCATACAACTTTCTCTCCTTA
901 Left GTAGGGGCTGAATGTTATCACAGC
902 Right TACAACTTTCTCTCCTTAAGCCTCA 917 Left GCAGTGTAGGGGCTGAATGTTAT
918 Right ACAACTTTCTCTCCTTAAGCCTCA
903 Left GAGGACAAGCCTTGACATTCAG
904 Right TACAACTTTCTCTCCTTAAGCCTCA 919 Left ATGTTGGCTTACATTAACTCCCATA
__________________________________________ 920 Right CAAAGTCCCTCTCCTTTGCAT
ALK Exon24 130-150 bases Seq. . 939 Left AGTGGCCCGCTTCTGTCT
Primer Sequence ID 940 Right ATGACAGGAAGAGCACAGTCAC
921 Left AGTGGCCCGCTTCTGTCT
922 Right GATGACAGGAAGAGCACAGTCAC Seq. .
ID Pnmer Sequence 923 Left CGCTTCTGTCTCCCCACAG 941 Left CGCTTCTGTCTCCCCACAG
924 Right GATGACAGGAAGAGCACAGTCAC 942 Right ATGACAGGAAGAGCACAGTCAC
925 Left CGCTTCTGTCTCCCCACA 943 Left CGCTTCTGTCTCCCCACA
926 Right GATGACAGGAAGAGCACAGTCAC 944 Right ATGACAGGAAGAGCACAGTCAC
927 Left AGTGGCCCGCTTCTGTCT 945 Left AGTGGCCCGCTTCTGTCT
928 Right AGGATGACAGGAAGAGCACAGT 946 Right AGGATGACAGGAAGAGCACAGTC
929 Left CGCTTCTGTCTCCCCACAG 947 Left CGCTTCTGTCTCCCCACAG
930 Right AGGATGACAGGAAGAGCACAGT 948 Right AGGATGACAGGAAGAGCACAGTC
931 Left CGCTTCTGTCTCCCCACA 949 Left CGCTTCTGTCTCCCCACA
932 Right AGGATGACAGGAAGAGCACAGT 950 Right AGGATGACAGGAAGAGCACAGTC
933 Left AGTGGCCCGCTTCTGTCT 951 Left AGTGGCCCGCTTCTGTCT
934 Right GATGACAGGAAGAGCACAGTCA 952 Right GGATGACAGGAAGAGCACAGTC
935 Left CGCTTCTGTCTCCCCACAG 953 Left CGCTTCTGTCTCCCCACAG
936 Right GATGACAGGAAGAGCACAGTCA 954 Right GGATGACAGGAAGAGCACAGTC
937 Left CGCTTCTGTCTCCCCACA 955 Left CGCTTCTGTCTCCCCACAG
938 Right GATGACAGGAAGAGCACAGTCA 956 Right GACAGGATGACAGGAAGAGCAC

957 Left CGCTTCTGTCTCCCCACA 959 Left CGCTTCTGTCTCCCCACA
958 Right GGATGACAGGAAGAGCACAGTC 960 Right GACAGGATGACAGGAAGAGCAC
ALK Exon24 161-200 bases S eq.
Primer Sequence ID 981 Left ATTTCAGATTTCCCTCCTCTCACT
961 Left ATTTCAGATTTCCCTCCTCTCACT 982 Right AGGATGACAGGAAGAGCACAGTC
962 Right GATGACAGGAAGAGCACAGTCAC
S eq. 983 Left ATTTCAGATTTCCCTCCTCTCACT
Primer Sequence ID 984 Right GGATGACAGGAAGAGCACAGTC
963 Left ATTTCAGATTTCCCTCCTCTCACT ________ 964 Right AGGATGACAGGAAGAGCACAGT 985 Left TTTCAGATTTCCCTCCTCTCACT
965 Left ATTTCAGATTTCCCTCCTCTCACT 986 Right GATGACAGGAAGAGCACAGTCA
966 Right GATGACAGGAAGAGCACAGTCA _________ __________________________________________ 987 Left ATTTCAGATTTCCCTCCTCTCAC
967 Left ATTTCAGATTTCCCTCCTCTCAC 988 Right AGGATGACAGGAAGAGCACAGTC
968 Right GATGACAGGAAGAGCACAGTCAC ________ __________________________________________ 989 Left TTTCAGATTTCCCTCCTCTCACT
969 Left ATTTCAGATTTCCCTCCTCTCAC 990 Right ATGACAGGAAGAGCACAGTCAC
970 Right AGGATGACAGGAAGAGCACAGT _________ __________________________________________ 991 Left ATTTCAGATTTCCCTCCTCTCAC
971 Left ATTTCAGATTTCCCTCCTCTCACT 992 Right GGATGACAGGAAGAGCACAGTC
972 Right ATGACAGGAAGAGCACAGTCAC
__________________________________________ 993 Left ATTTCAGATTTCCCTCCTCTCACT
973 Left ATTTCAGATTTCCCTCCTCTCAC 994 Right AGGATGACAGGAAGAGCACAG
974 Right GATGACAGGAAGAGCACAGTCA _________ __________________________________________ 995 Left ATTTCCCTCCTCTCACTGACAA
975 Left ATTTCAGATTTCCCTCCTCTCAC 996 Right GATGACAGGAAGAGCACAGTCAC
976 Right ATGACAGGAAGAGCACAGTCAC
__________________________________________ 997 Left ATTTCCCTCCTCTCACTGACAA
977 Left TTTCAGATTTCCCTCCTCTCACT 998 Right AGGATGACAGGAAGAGCACAGT
978 Right GATGACAGGAAGAGCACAGTCAC ________ __________________________________________ 999 Left CATTTCAGATTTCCCTCCTCTCACT
979 Left TTTCAGATTTCCCTCCTCTCACT 1000 Right GATGACAGGAAGAGCACAGTCAC

980 Right AGGATGACAGGAAGAGCACAGT
ALK Exon24 201-300 bases Sea ID' Primer Sequence _1011 Left ATTTCAGATTTCCCTCCTCTCACT
1001 Left ATTTCAGATTTCCCTCCTCTCACT 1012 Right AGGGAGACCTAGTATTCTGCTCTGA
1002 Right GAGACCTAGTATTCTGCTCTGAAGG _ ________________________________________________ 1013 Left ATTTCAGATTTCCCTCCTCTCACT
1003 Left ATTTCAGATTTCCCTCCTCTCACT 1014 Right TCTGGAGGGAGACCTAGTATTCTG
1004 Right GGAGACCTAGTATTCTGCTCTGAAG
1015 Left ATTTCAGATTTCCCTCCTCTCAC
1005 Left ATTTCAGATTTCCCTCCTCTCACT _1016 Right CTCTGGAGGGAGACCTAGTATTCTG
1006 Right CTCTGGAGGGAGACCTAGTATTCTG Seq.
ID Primer Sequence 1007 Left ATTTCAGATTTCCCTCCTCTCAC 1017 Left ATTTCAGATTTCCCTCCTCTCACT
1008 Right GAGACCTAGTATTCTGCTCTGAAGG 1018 Right GGGAGACCTAGTATTCTGCTCTGA
1009 Left ATTTCAGATTTCCCTCCTCTCAC 1019 Left ATTTCAGATTTCCCTCCTCTCAC
1010 Right GGAGACCTAGTATTCTGCTCTGAAG 1020 Right AGGGAGACCTAGTATTCTGCTCTGA

1021 Left ATTTCAGATTTCCCTCCTCTCAC 1031 Left TTTCAGATTTCCCTCCTCTCACT
1022 Right TCTGGAGGGAGACCTAGTATTCTG 1032 Right CTCTGGAGGGAGACCTAGTATTCTG
1023 Left CTCCTCTCACTGACAAGCTCCT 1033 Left TTTCAGATTTCCCTCCTCTCACT
1024 Right AAACAAAGCTGAATCATCCTACATC 1034 Right AGGGAGACCTAGTATTCTGCTCTGA
1025 Left ATTTCAGATTTCCCTCCTCTCAC 1035 Left TTTCAGATTTCCCTCCTCTCACT
1026 Right GGGAGACCTAGTATTCTGCTCTGA 1036 Right TCTGGAGGGAGACCTAGTATTCTG
1027 Left TTTCAGATTTCCCTCCTCTCACT 1037 Left ATTTCAGATTTCCCTCCTCTCACT
1028 Right GAGACCTAGTATTCTGCTCTGAAGG 1038 Right GCTCTGGAGGGAGACCTAGTATTC
1029 Left TTTCAGATTTCCCTCCTCTCACT 1039 Left TTTCAGATTTCCCTCCTCTCACT
1030 Right GGAGACCTAGTATTCTGCTCTGAAG 1040 Right GGGAGACCTAGTATTCTGCTCTGA
ALK Exon24 301-400 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
1041 Left ATTTCAGATTTCCCTCCTCTCACT 1061 Left ATTTCAGATTTCCCTCCTCTCACT
1042 Right AAACAAAGCTGAATCATCCTACATC 1062 Right CAATAAAACAAAGCTGAATCATCCTA
1043 Left ATTTCAGATTTCCCTCCTCTCACT 1063 Left ATTTCAGATTTCCCTCCTCTCACT
1044 Right ACAATAAAACAAAGCTGAATCATCC 1064 Right AGCTGAATCATCCTACATCCAAAT
1045 Left ATTTCAGATTTCCCTCCTCTCAC 1065 Left AATGCCTCCAGGTGATTTCTAAT
1046 Right AAACAAAGCTGAATCATCCTACATC 1066 Right GAGACCTAGTATTCTGCTCTGAAGG
1047 Left ATTTCAGATTTCCCTCCTCTCAC 1067 Left AATGCCTCCAGGTGATTTCTAAT
1048 Right ACAATAAAACAAAGCTGAATCATCC 1068 Right GGAGACCTAGTATTCTGCTCTGAAG
1049 Left ATTTCAGATTTCCCTCCTCTCACT 1069 Left TTTCAGATTTCCCTCCTCTCACT
1050 Right AAAACAAAGCTGAATCATCCTACAT 1070 Right AAAACAAAGCTGAATCATCCTACAT
1051 Left ATTTCAGATTTCCCTCCTCTCAC 1071 Left ATTTCAGATTTCCCTCCTCTCAC
1052 Right AAAACAAAGCTGAATCATCCTACAT 1072 Right CAATAAAACAAAGCTGAATCATCCTA
1053 Left ATTTCAGATTTCCCTCCTCTCACT 1073 Left ATTTCAGATTTCCCTCCTCTCACT
1054 Right TAAAACAAAGCTGAATCATCCTACA 1074 Right AAGCTGAATCATCCTACATCCAAAT
1055 Left TTTCAGATTTCCCTCCTCTCACT 1075 Left AAATGCCTCCAGGTGATTTCTAAT
1056 Right AAACAAAGCTGAATCATCCTACATC 1076 Right GAGACCTAGTATTCTGCTCTGAAGG
1057 Left ATTTCAGATTTCCCTCCTCTCAC 1077 Left AAATGCCTCCAGGTGATTTCTAAT
1058 Right TAAAACAAAGCTGAATCATCCTACA 1078 Right GGAGACCTAGTATTCTGCTCTGAAG
1059 Left TTTCAGATTTCCCTCCTCTCACT 1079 Left AATGCCTCCAGGTGATTTCTAAT
1060 Right ACAATAAAACAAAGCTGAATCATCC 1080 Right CTCTGGAGGGAGACCTAGTATTCTG
ALK Exon24 401-600 bases Seq. 082 Right AAACAAAGCTGAATCATCCTACATC
ID Primer Sequence 1081 Left TGCACAATAAATTAAAAGGGAAAGA 083 Left TGCACAATAAATTAAAAGGGAAAGA

1084 Right ACAATAAAACAAAGCTGAATCATCC
1103 Left TGCACAATAAATTAAAAGGGAAAGA
1085 Left GTGCACAATAAATTAAAAGGGAAAG 1104 Right GAGACCTAGTATTCTGCTCTGAAGG
1086 Right AAACAAAGCTGAATCATCCTACATC
1105 Left TGCACAATAAATTAAAAGGGAAAGA
1087 Left TGCACAATAAATTAAAAGGGAAAGA 1106 Right GGAGACCTAGTATTCTGCTCTGAAG
1088 Right AAAACAAAGCTGAATCATCCTACAT
S eq. 1107 Left TGTGCACAATAAATTAAAAGGGAAA
Primer Sequence ID 1108 Right ACAATAAAACAAAGCTGAATCATCC
1089 Left ATCATATTACCTGGGAAGACTTCAA ______ ¨
1090 Right AAACAAAGCTGAATCATCCTACATC 1109 Left TGTTTATAAATTGGGGGTATTCAAA
1110 Right GAGACCTAGTATTCTGCTCTGAAGG
1091 Left GTGCACAATAAATTAAAAGGGAAAG
1092 Right ACAATAAAACAAAGCTGAATCATCC 1111 Left TTGTTTATAAATTGGGGGTATTCAA
1112 Right GAGACCTAGTATTCTGCTCTGAAGG
1093 Left TGTGCACAATAAATTAAAAGGGAAA
1094 Right AAACAAAGCTGAATCATCCTACATC 1113 Left TGTTTATAAATTGGGGGTATTCAAA
1114 Right GGAGACCTAGTATTCTGCTCTGAAG
1095 Left TGCACAATAAATTAAAAGGGAAAGA
1096 Right TAAAACAAAGCTGAATCATCCTACA 1115 Left TTGTTTATAAATTGGGGGTATTCAA
1116 Right GGAGACCTAGTATTCTGCTCTGAAG
1097 Left TAAATTAAAAGGGAAAGAACACCTG
1098 Right AAACAAAGCTGAATCATCCTACATC 1117 Le ft GTGCACAATAAATTAAAAGGGAAAG
1118 Right AAAACAAAGCTGAATCATCCTACAT
1099 Left GCACAATAAATTAAAAGGGAAAGAA
1100 Right AAACAAAGCTGAATCATCCTACATC 1119 Left TAAATTAAAAGGGAAAGAACACCTG
¨1120 Right ACAATAAAACAAAGCTGAATCATCC
1101 Left TGGGAAGACTTCAAATGTACAAATA
1102 Right AAACAAAGCTGAATCATCCTACATC
ALK Exon24 601-800 bases Seq.
Primer Sequence ID 1135 Left CTTGTTTATAAATTGGGGGTATTCA
1121 Left TGTTTATAAATTGGGGGTATTCAAA 1136 Right AAACAAAGCTGAATCATCCTACATC
1122 Right AAACAAAGCTGAATCATCCTACATC
1137 Left TGCACAATAAATTAAAAGGGAAAGA
1123 Left TTGTTTATAAATTGGGGGTATTCAA 1138 Right GCAAGTGAATCCCTGATAGAATAAG
1124 Right AAACAAAGCTGAATCATCCTACATC
1139 Left CTGGATCTGCTTGAAGAAAATTAGT
1125 Left TGTTTATAAATTGGGGGTATTCAAA ¨1140 Right AAACAAAGCTGAATCATCCTACATC
1126 Right ACAATAAAACAAAGCTGAATCATCC
1127 Left TGCACAATAAATTAAAAGGGAAAGA S eq.
1128 Right AGTTACCATCTCAAAGACAAAGCTG ID Primer Sequence __________________________________________ 1141 Left CAAACTTGTTTATAAATTGGGGGTA
1129 Left TGTTTATAAATTGGGGGTATTCAAA ¨1142 Right ACAATAAAACAAAGCTGAATCATCC
1130 Right AGTTACCATCTCAAAGACAAAGCTG ______ __________________________________________ 1143 Left TTTATAAATTGGGGGTATTCAAATG
1131 Left TTGTTTATAAATTGGGGGTATTCAA ¨1144 Right AAACAAAGCTGAATCATCCTACATC
1132 Right AGTTACCATCTCAAAGACAAAGCTG ____ _________________________________________ ¨1145 Left TGTTTATAAATTGGGGGTATTCAAA

1133 Left CAAACTTGTTTATAAATTGGGGGTA ¨1146 Right AAAACAAAGCTGAATCATCCTACAT
1134 Right AAACAAAGCTGAATCATCCTACATC ______ 1147 Left TTGTTTATAAATTGGGGGTATTCAA 154 Right GCGAGGATATTTTATGACACTTGTT
1148 Right AAAACAAAGCTGAATCATCCTACAT
155 Left TTTATAAATTGGGGGTATTCAAATG
1149 Left CTGGATCTGCTTGAAGAAAATTAGT 156 Right ACAATAAAACAAAGCTGAATCATCC
1150 Right ACAATAAAACAAAGCTGAATCATCC
157 Left ATCTCCTTTTGAATGAAAGAGACCT
1151 Left GTGCACAATAAATTAAAAGGGAAAG 158 Right GAGACCTAGTATTCTGCTCTGAAGG
1152 Right AGTTACCATCTCAAAGACAAAGCTG
159 Left ATCTCCTTTTGAATGAAAGAGACCT
1153 Left ATCATATTACCTGGGAAGACTTCAA 160 Right GGAGACCTAGTATTCTGCTCTGAAG
ALK Exon24 801-1000 bases Seq.
Primer Sequence Primer Sequence ID ID
1161 Left TAGGAATTAAAAGAGAGGCCAAGAT 1181 Left TTTAGGAATTAAAAGAGAGGCCAAG
1162 Right AAACAAAGCTGAATCATCCTACATC 1182 Right AAACAAAGCTGAATCATCCTACATC
1163 Left ATCTCCTTTTGAATGAAAGAGACCT 1183 Left TAGGAATTAAAAGAGAGGCCAAGAT
1164 Right AAACAAAGCTGAATCATCCTACATC 1184 Right AAAACAAAGCTGAATCATCCTACAT
1165 Left TAGGAATTAAAAGAGAGGCCAAGAT 1185 Left ATCTCCTTTTGAATGAAAGAGACCT
1166 Right ACAATAAAACAAAGCTGAATCATCC 1186 Right AAAACAAAGCTGAATCATCCTACAT
1167 Left ATCTCCTTTTGAATGAAAGAGACCT 1187 Left AAAGCTTGAGATAGCTCATAATTGC
1168 Right ACAATAAAACAAAGCTGAATCATCC 1188 Right AAACAAAGCTGAATCATCCTACATC
1169 Left TGCACAATAAATTAAAAGGGAAAGA 1189 Left TTTAGGAATTAAAAGAGAGGCCAAG
1170 Right AAAGATGACTAAAACAGCATCCTTG 1190 Right ACAATAAAACAAAGCTGAATCATCC
1171 Left ACGTCAGGGATTTAGGAATTAAAAG 1191 Left ACGTCAGGGATTTAGGAATTAAAAG
1172 Right ACAATAAAACAAAGCTGAATCATCC 1192 Right AAAACAAAGCTGAATCATCCTACAT
1173 Left TGCACAATAAATTAAAAGGGAAAGA 1193 Left TGTTTATAAATTGGGGGTATTCAAA
1174 Right GCGAGGATATTTTATGACACTTGTT 1194 Right GCAAGTGAATCCCTGATAGAATAAG
1175 Left TGTTTATAAATTGGGGGTATTCAAA 1195 Left TTGTTTATAAATTGGGGGTATTCAA
1176 Right GCGAGGATATTTTATGACACTTGTT 1196 Right GCAAGTGAATCCCTGATAGAATAAG
1177 Left TTGTTTATAAATTGGGGGTATTCAA 1197 Left GTGCACAATAAATTAAAAGGGAAAG
1178 Right GCGAGGATATTTTATGACACTTGTT 1198 Right AAAGATGACTAAAACAGCATCCTTG
1179 Left TGCACAATAAATTAAAAGGGAAAGA 1199 Left CAAACTTGTTTATAAATTGGGGGTA
1180 Right TTGTCAAAAATGCAATTCCTTAACT 1200 Right GCGAGGATATTTTATGACACTTGTT
ALK Exon24 2kb S eq. . 1205 Left ATCTCCTTTTGAATGAAAGAGACCT
Pnmer S equence ID 1206 Right CTAAAAGGATGAAGTGACAGGAAGA
1201 Left ATCTCCTTTTGAATGAAAGAGACCT
1202 Right CATGTTAGGAGTGACTTTGGAACTT 1207 Left ATCTCCTTTTGAATGAAAGAGACCT
1208 Right TAAAAGGATGAAGTGACAGGAAGAG
1203 Left ACTGTAGTCACATACATACGCTCCA
1204 Right AAACAAAGCTGAATCATCCTACATC 1209 Left ACTGTAGTCACATACATACGCTC CA
1210 Right GCGAGGATATTTTATGACACTTGTT

224 Right ATAAGCTCTTCTGAGAGTTGACTGC
__________________________________________ 1225 Left AGGCACTCTCTCTTCCATTTTAACT
S eq. . 1226 Right AAACAAAGCTGAATCATCCTACATC
ID Primer Sequence 1211 Left ACTGTAGTCACATACATACGCTCCA 1227 Left TGCACAATAAATTAAAAGGGAAAGA
1212 Right ACAATAAAACAAAGCTGAATCATCC 1228 Right AAGGGCTGAAAAACTACCTTAAAAA
1213 Left ACGTCAGGGATTTAGGAATTAAAAG 1229 Left TGCACAATAAATTAAAAGGGAAAGA
1214 Right CAAGTGTACTTCCTGACCTCTCATT 1230 Right AAAGGGCTGAAAAACTACCTTAAAA
1215 Left TGCACAATAAATTAAAAGGGAAAGA 1231 Left TGCACAATAAATTAAAAGGGAAAGA
1216 Right CATGTTAGGAGTGACTTTGGAACTT 1232 Right TTAGAAGGAGCAGATGGTAAAGCTA
1217 Left ACTGTAGTCACATACATACGCTCCA 1233 Left TGTTTATAAATTGGGGGTATTCAAA
1218 Right AGTTACCATCTCAAAGACAAAGCTG 1234 Right AAGGGCTGAAAAACTACCTTAAAAA
1219 Left TGTTTATAAATTGGGGGTATTCAAA 1235 Left TGTTTATAAATTGGGGGTATTCAAA
1220 Right CATGTTAGGAGTGACTTTGGAACTT 1236 Right AAAGGGCTGAAAAACTACCTTAAAA
1237 Left TTGTTTATAAATTGGGGGTATTCAA
1238 Right AAAGGGCTGAAAAACTACCTTAAAA
1221 Left TTGTTTATAAATTGGGGGTATTCAA ______ 1222 Right CATGTTAGGAGTGACTTTGGAACTT 1239 Left TTGTTTATAAATTGGGGGTATTCAA
1240 Right AAGGGCTGAAAAACTACCTTAAAAA
1223 Left ATCTCCTTTTGAATGAAAGAGACCT
ALK Exon25 130-160 bases Seq. .
Pnmer Sequence ID 1257 Left TCCTAGGGATAAAATTAGGAAATGC
1241 Left TCCTAGGGATAAAATTAGGAAATGC 1258 Right GAGGGGTGAGGCAGTCTTTA
1242 Right AGGGGTGAGGCAGTCTTTACTC
1259 Left TTCCTAGGGATAAAATTAGGAAATG
1243 Left TCCTAGGGATAAAATTAGGAAATGC 1260 Right AGGGGTGAGGCAGTCTTTACTC
1244 Right GAGGGGTGAGGCAGTCTTTACT
1245 Left TCCTAGGGATAAAATTAGGAAATGC
1246 Right GGGGTGAGGCAGTCTTTACTC
__________________________________________ S eq. .
1247 Left TCCTAGGGATAAAATTAGGAAATGC ID Primer Sequence 1248 Right GAGGGGTGAGGCAGTCTTTAC 1261 Left TCCTAGGGATAAAATTAGGAAATGC
1262 Right GGGGTGAGGCAGTCTTTACTCA
1249 Left TCCTAGGGATAAAATTAGGAAATGC _______ 1250 Right GGGTGAGGCAGTCTTTACTCA 1263 Left TTCCTAGGGATAAAATTAGGAAATG
1264 Right GGGGTGAGGCAGTCTTTACTC
1251 Left TCCTAGGGATAAAATTAGGAAATGC _______ 1252 Right GAGGGGTGAGGCAGTCTTTACTC 1265 Left TTCCTAGGGATAAAATTAGGAAATGC
1266 Right AGGGGTGAGGCAGTCTTTACTC
1253 Left TCCTAGGGATAAAATTAGGAAATGC _______ 1254 Right AGGGGTGAGGCAGTCTTTACT 1267 Left CTTCCTAGGGATAAAATTAGGAAATG
1268 Right GGGGTGAGGCAGTCTTTACTC
1255 Left TCCTAGGGATAAAATTAGGAAATGC _______ 1256 Right AGGGGTGAGGCAGTCTTTACTCA 1269 Left TCCTAGGGATAAAATTAGGAAATGC

ALK Exon25 161-200 bases 1270 Right GAGGGGTGAGGCAGTCTTTACTCA 275 Left CCTAGGGATAAAATTAGGAAATGC
276 Right GAGGGGTGAGGCAGTCTTTACT
1271 Left TTCCTAGGGATAAAATTAGGAAATGC
1272 Right GGGGTGAGGCAGTCTTTACTC
277 Left TTCCTAGGGATAAAATTAGGAAATG
278 Right GGGTGAGGCAGTCTTTACTCA
1273 Left CCTAGGGATAAAATTAGGAAATGC
1274 Right AGGGGTGAGGCAGTCTTTACTC
279 Left TTCCTAGGGATAAAATTAGGAAATG
280 Right AGGGGTGAGGCAGTCTTTACT
Seq. ID
Primer Sequence ID
1301 Left TCATCTTCCTAGGGATAAAATTAGG
1281 Left CTTGGAGATAAAATCCTAGTGATGG ¨1302 Right GAGGGGTGAGGCAGTCTTTAC
1282 Right GGGGTGAGGCAGTCTTTACTC
1303 Left TTCCTAGGGATAAAATTAGGAAATG
1283 Left TCCTAGGGATAAAATTAGGAAATGC 1304 Right GAGGGGTGAGGCAGTCTTTAC
1284 Right GCTGAGGTGGAAGAGACAGG
1305 Left CTCATCTTCCTAGGGATAAAATTAGG
1285 Left TCCTAGGGATAAAATTAGGAAATGC 1306 Right AGGGGTGAGGCAGTCTTTACTC
1286 Right GGCTGAGGTGGAAGAGACAG
1307 Left CTCATCTTCCTAGGGATAAAATTAGG
1287 Left CTTGGAGATAAAATCCTAGTGATGG 1308 Right GAGGGGTGAGGCAGTCTTTACT
1288 Right GGGTGAGGCAGTCTTTACTCA
¨1309 Left CTTCCTAGGGATAAAATTAGGAAATG
1289 Left TGTACACTCATCTTCCTAGGGATAAA 1310 Right AGGGGTGAGGCAGTCTTTACTC
1290 Right GAGGGGTGAGGCAGTCTTTACT
1311 Left CTTCCTAGGGATAAAATTAGGAAATG
1291 Left TGTACACTCATCTTCCTAGGGATAAA 312 Right GAGGGGTGAGGCAGTCTTTACT
1292 Right GAGGGGTGAGGCAGTCTTTAC
1313 Left CTCATCTTCCTAGGGATAAAATTAGG
1293 Left TCATCTTCCTAGGGATAAAATTAGG ¨1314 Right GGGGTGAGGCAGTCTTTACTC
1294 Right AGGGGTGAGGCAGTCTTTACTC
1315 Left CTCATCTTCCTAGGGATAAAATTAGG
1295 Left TCATCTTCCTAGGGATAAAATTAGG 1316 Right GAGGGGTGAGGCAGTCTTTAC
1296 Right GAGGGGTGAGGCAGTCTTTACT
¨1317 Left TTCCTAGGGATAAAATTAGGAAATGC
1297 Left TTCCTAGGGATAAAATTAGGAAATG 1318 Right GAGGGGTGAGGCAGTCTTTACT
1298 Right GAGGGGTGAGGCAGTCTTTACT
1319 Left CTTCCTAGGGATAAAATTAGGAAATG
1299 Left TCATCTTCCTAGGGATAAAATTAGG 1320 Right GAGGGGTGAGGCAGTCTTTAC
1300 Right GGGGTGAGGCAGTCTTTACTC
Seq. Primer Sequence ALK Exon25 201-300 bases Seq.
1325 Left TCCTAGGGATAAAATTAGGAAATGC
Primer Sequence ID
1326 Right CATAGCCTGAAAAGGAACTTAGTGA
1321 Left TCCTAGGGATAAAATTAGGAAATGC
1322 Right AGCCTGAAAAGGAACTTAGTGAAAT 1327 Left TCCTAGGGATAAAATTAGGAAATGC
__________________________________________ 1328 Right CTAATTAAGGTTTCCCATAGCCTGA
1323 Left TCCTAGGGATAAAATTAGGAAATGC
1324 Right TAGCCTGAAAAGGAACTTAGTGAAA 1329 Left TCCTAGGGATAAAATTAGGAAATGC
1330 Right ATAGCCTGAAAAGGAACTTAGTGAAA

1345 Left TGTACACTCATCTTCCTAGGGATAAA
1331 Left TCCTAGGGATAAAATTAGGAAATGC 1346 Right TAGCCTGAAAAGGAACTTAGTGAAA
1332 Right TAGCCTGAAAAGGAACTTAGTGAAAT
Seq. . 1347 Left TCCTAGGGATAAAATTAGGAAATGC
Pnmer Sequence ID 1348 Right CCATAGCCTGAAAAGGAACTTAGTG
1333 Left TCCTAGGGATAAAATTAGGAAATGC
1334 Right AGGTAGAAAGTTGACAGGGTACCAG 1349 Left CTTGGAGATAAAATCCTAGTGATGG
__________________________________________ 1350 Right AGGTAGAAAGTTGACAGGGTACCAG
1335 Left TCCTAGGGATAAAATTAGGAAATGC
1336 Right AGCCTGAAAAGGAACTTAGTGAAA 1351 Left TGTACACTCATCTTCCTAGGGATAAA
1352 Right CATAGCCTGAAAAGGAACTTAGTGA
1337 Left TCCTAGGGATAAAATTAGGAAATGC
1338 Right TAATTAAGGTTTCCCATAGCCTGA 1353 Left TCCTAGGGATAAAATTAGGAAATGC
1354 Right CACTAATTAAGGTTTCCCATAGCC
1339 Left TCCTAGGGATAAAATTAGGAAATGC
1340 Right TAATTAAGGTTTCCCATAGCCTGAA 1355 Left TCATCTTCCTAGGGATAAAATTAGG
1356 Right AGCCTGAAAAGGAACTTAGTGAAAT
1341 Left TCCTAGGGATAAAATTAGGAAATGC
1342 Right GGTAGAAAGTTGACAGGGTACCAG 1357 Left TCCTAGGGATAAAATTAGGAAATGC
1358 Right GCCTGAAAAGGAACTTAGTGAAAT
1343 Left TGTACACTCATCTTCCTAGGGATAAA
1344 Right AGCCTGAAAAGGAACTTAGTGAAAT 1359 Left AACTTCAGCTTGGAGATAAAATCCT
__________________________________________ 1360 Right ACAGGGTACCAGGAGATGATGTAAG
ALK Exon25 301-400 bases Seq. . 1379 Left AAACTTCAGCTTGGAGATAAAATCC
Primer Sequence ID 1380 Right TGGTCACTAATTAAGGTTTCCCATA
1361 Left AACTTCAGCTTGGAGATAAAATCCT
1362 Right TGGTCACTAATTAAGGTTTCCCATA Seq. .
ID Primer Sequence 1363 Left GGGAAGGAACTATTTAAACTTCAGC 1381 Left AAACTTCAGCTTGGAGATAAAATCC
1364 Right TGGTCACTAATTAAGGTTTCCCATA 1382 Right AGCCTGAAAAGGAACTTAGTGAAAT
1365 Left TCCTAGGGATAAAATTAGGAAATGC 1383 Left AAACTTCAGCTTGGAGATAAAATCC
1366 Right TGGTCACTAATTAAGGTTTCCCATA 1384 Right TAGCCTGAAAAGGAACTTAGTGAAA
1367 Left AACTTCAGCTTGGAGATAAAATCCT 1385 Left AAACTTCAGCTTGGAGATAAAATCC
1368 Right AGCCTGAAAAGGAACTTAGTGAAAT 1386 Right CATAGCCTGAAAAGGAACTTAGTGA
1369 Left GGGAAGGAACTATTTAAACTTCAGC 1387 Left CTTGGAGATAAAATCCTAGTGATGG
1370 Right AGCCTGAAAAGGAACTTAGTGAAAT 1388 Right TGGTCACTAATTAAGGTTTCCCATA
1371 Left AACTTCAGCTTGGAGATAAAATCCT 1389 Left CTTGGAGATAAAATCCTAGTGATGG
1372 Right TAGCCTGAAAAGGAACTTAGTGAAA 1390 Right AGCCTGAAAAGGAACTTAGTGAAAT
1373 Left GGGAAGGAACTATTTAAACTTCAGC 1391 Left CTTGGAGATAAAATCCTAGTGATGG
1374 Right TAGCCTGAAAAGGAACTTAGTGAAA 1392 Right TAGCCTGAAAAGGAACTTAGTGAAA
1375 Left AACTTCAGCTTGGAGATAAAATCCT 1393 Left GGAACTATTTAAACTTCAGCTTGGA
1376 Right CATAGCCTGAAAAGGAACTTAGTGA 1394 Right TGGTCACTAATTAAGGTTTCCCATA
1377 Left GGGAAGGAACTATTTAAACTTCAGC 1395 Left CTTGGAGATAAAATCCTAGTGATGG
1378 Right CATAGCCTGAAAAGGAACTTAGTGA 1396 Right CATAGCCTGAAAAGGAACTTAGTGA

1397 Left GGAACTATTTAAACTTCAGCTTGGA 1399 Left GGAACTATTTAAACTTCAGCTTGGA
1398 Right AGCCTGAAAAGGAACTTAGTGAAAT 1400 Right TAGCCTGAAAAGGAACTTAGTGAAA
ALK Exon25 401-600 bases S eq.
Primer S equence ID 1421 Left TTCCATTTCTCTCTTAGTTGTGAGG
1401 Left TTAATCATTTCCCCTAATCCTTTTC 1422 Right TGGTCACTAATTAAGGTTTCCCATA
1402 Right TGGTCACTAATTAAGGTTTCCCATA
1423 Left GAAGCACTTACAACAACACTTAGCA
1403 Left GAAGCACTTACAACAACACTTAGCA 1424 Right TAGCCTGAAAAGGAACTTAGTGAAA
1404 Right TGGTCACTAATTAAGGTTTCCCATA
S eq.1425 Left TGAAGCACTTACAACAACACTTAGC
Primer S equence ID 1426 Right TAGCCTGAAAAGGAACTTAGTGAAA
1405 Left TGAAGCACTTACAACAACACTTAGC _______ 1406 Right TGGTCACTAATTAAGGTTTCCCATA 1427 Left GAAGCACTTACAACAACACTTAGCA
1428 Right CATAGCCTGAAAAGGAACTTAGTGA
1407 Left ATTGTTAAGGCTGTTTCTCTCACAC ______ 1408 Right TGGTCACTAATTAAGGTTTCCCATA 1429 Left TGAAGCACTTACAACAACACTTAGC
1430 Right CATAGCCTGAAAAGGAACTTAGTGA
1409 Left GAAGCACTTACAACAACACTTAGCA _______ 1410 Right AGCCTGAAAAGGAACTTAGTGAAAT 1431 Left ATTGTTAAGGCTGTTTCTCTCACAC
1432 Right TAGCCTGAAAAGGAACTTAGTGAAA
1411 Left TGAAGCACTTACAACAACACTTAGC ______ 1412 Right AGCCTGAAAAGGAACTTAGTGAAAT 1433 Left GCACCTTGTGTCTTATAAGGTTGTT
1434 Right TAGCCTGAAAAGGAACTTAGTGAAA
1413 Left CATTTTTAATCATTTCCCCTAATCC ______ 1414 Right TGGTCACTAATTAAGGTTTCCCATA 1435 Left GAATTGTTAAGGCTGTTTCTCTCAC
1436 Right TGGTCACTAATTAAGGTTTCCCATA
1415 Left AGAATTGTTAAGGCTGTTTCTCTCA ______ 1416 Right TGGTCACTAATTAAGGTTTCCCATA 1437 Left ATTGTTAAGGCTGTTTCTCTCACAC
1438 Right CATAGCCTGAAAAGGAACTTAGTGA
1417 Left ATTGTTAAGGCTGTTTCTCTCACAC ______ 1418 Right AGCCTGAAAAGGAACTTAGTGAAAT 1439 Left GCACCTTGTGTCTTATAAGGTTGTT
1440 Right CATAGCCTGAAAAGGAACTTAGTGA
1419 Left GCACCTTGTGTCTTATAAGGTTGTT
1420 Right AGCCTGAAAAGGAACTTAGTGAAAT
ALK Exon25 401-600 bases S eq. 1450 Right AGCCTGAAAAGGAACTTAGTGAAAT
ID Primer Sequence 1441 Left TTAATCATTTCCCCTAATCCTTTTC 1451 Left TGAAGCACTTACAACAACACTTAGC
1442 Right TGGTCACTAATTAAGGTTTCCCATA 1452 Right AGCCTGAAAAGGAACTTAGTGAAAT
1443 Left GAAGCACTTACAACAACACTTAGCA ¨1453 Left CATTTTTAATCATTTCCCCTAATCC
1444 Right TGGTCACTAATTAAGGTTTCCCATA 1454 Right TGGTCACTAATTAAGGTTTCCCATA
1445 Left TGAAGCACTTACAACAACACTTAGC 1455 Left AGAATTGTTAAGGCTGTTTCTCTCA
1446 Right TGGTCACTAATTAAGGTTTCCCATA 1456 Right TGGTCACTAATTAAGGTTTCCCATA
_ 1447 Left ATTGTTAAGGCTGTTTCTCTCACAC 1457 Left ATTGTTAAGGCTGTTTCTCTCACAC
1448 Right TGGTCACTAATTAAGGTTTCCCATA ¨1458 Right AGCCTGAAAAGGAACTTAGTGAAAT
__________________________________________ Seq.
Primer Sequence 1449 Left GAAGCACTTACAACAACACTTAGCA ID

1459 Left GCACCTTGTGTCTTATAAGGTTGTT 1470 Right CATAGCCTGAAAAGGAACTTAGTGA
1460 Right AGCCTGAAAAGGAACTTAGTGAAAT
1471 Left ATTGTTAAGGCTGTTTCTCTCACAC
1461 Left TTCCATTTCTCTCTTAGTTGTGAGG 1472 Right TAGCCTGAAAAGGAACTTAGTGAAA
1462 Right TGGTCACTAATTAAGGTTTCCCATA
1473 Left GCACCTTGTGTCTTATAAGGTTGTT
1463 Left GAAGCACTTACAACAACACTTAGCA 1474 Right TAGCCTGAAAAGGAACTTAGTGAAA
1464 Right TAGCCTGAAAAGGAACTTAGTGAAA
1475 Left GAATTGTTAAGGCTGTTTCTCTCAC
1465 Left TGAAGCACTTACAACAACACTTAGC 1476 Right TGGTCACTAATTAAGGTTTCCCATA
1466 Right TAGCCTGAAAAGGAACTTAGTGAAA
1477 Left ATTGTTAAGGCTGTTTCTCTCACAC
1467 Left GAAGCACTTACAACAACACTTAGCA 1478 Right CATAGCCTGAAAAGGAACTTAGTGA
1468 Right CATAGCCTGAAAAGGAACTTAGTGA
1479 Left GCACCTTGTGTCTTATAAGGTTGTT
1469 Left TGAAGCACTTACAACAACACTTAGC 1480 Right CATAGCCTGAAAAGGAACTTAGTGA
ALK Exon25 601-800 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
1481 Left GGACAGTAATAGCACCTTGTGTCTT 1501 Left AAATGAGGACAGTAATAGCACCTTG
1482 Right TGGTCACTAATTAAGGTTTCCCATA 1502 Right CATAGCCTGAAAAGGAACTTAGTGA
1483 Left AAATGAGGACAGTAATAGCACCTTG 1503 Left TCCTAGGGATAAAATTAGGAAATGC
1484 Right TGGTCACTAATTAAGGTTTCCCATA 1504 Right AGTATCCAAGTTATCCCATGTCTCA
1485 Left GGACAGTAATAGCACCTTGTGTCTT 1505 Left TCCTAGGGATAAAATTAGGAAATGC
1486 Right AGCCTGAAAAGGAACTTAGTGAAAT 1506 Right GTATCCAAGTTATCCCATGTCTCAG
1487 Left AAATGAGGACAGTAATAGCACCTTG 1507 Left TCCTAGGGATAAAATTAGGAAATGC
1488 Right AGCCTGAAAAGGAACTTAGTGAAAT 1508 Right TCAATAAAGCTCACTTTGAAGGTCT
1489 Left GCACCTTGTGTCTTATAAGGTTGTT 1509 Left TCCTAGGGATAAAATTAGGAAATGC
1490 Right TGGTCACTAATTAAGGTTTCCCATA 1510 Right AGTTGAGGAAGTTCAATAAAGCTCA
1491 Left GGACAGTAATAGCACCTTGTGTCTT 1511 Left TCCTAGGGATAAAATTAGGAAATGC
1492 Right TAGCCTGAAAAGGAACTTAGTGAAA 1512 Right TTGAGGAAGTTCAATAAAGCTCACT
1493 Left AAATGAGGACAGTAATAGCACCTTG 1513 Left TCCTAGGGATAAAATTAGGAAATGC
1494 Right TAGCCTGAAAAGGAACTTAGTGAAA 1514 Right TGAGGAAGTTCAATAAAGCTCACTT
1495 Left AACTTCAGCTTGGAGATAAAATCCT 1515 Left TCCTAGGGATAAAATTAGGAAATGC
1496 Right AGTATCCAAGTTATCCCATGTCTCA 1516 Right CTGAGGAGTTGAGGAAGTTCAATAA
1497 Left AACTTCAGCTTGGAGATAAAATCCT 1517 Left TCCTAGGGATAAAATTAGGAAATGC
1498 Right GTATCCAAGTTATCCCATGTCTCAG 1518 Right GGAAGTTCAATAAAGCTCACTTTGA
1499 Left GGACAGTAATAGCACCTTGTGTCTT 1519 Left TCCTAGGGATAAAATTAGGAAATGC
1500 Right CATAGCCTGAAAAGGAACTTAGTGA 1520 Right TTCAATAAAGCTCACTTTGAAGGTC
ALK Exon25 801-1000 bases Seq.1521 Left AACTTCAGCTTGGAGATAAAATCCT
Primer Sequence ID 1522 Right AGTACTGAGGAGTTGAGGAAGTTCA

1542 Right TCAATAAAGCTCACTTTGAAGGTCT
1523 Left TTAATCATTTCCCCTAATCCTTTTC
1524 Right AGTACTGAGGAGTTGAGGAAGTTCA 1543 Left TTAATCATTTCCCCTAATCCTTTTC
1544 Right AGTTGAGGAAGTTCAATAAAGCTCA
1525 Left TTAATCATTTCCCCTAATCCTTTTC
1526 Right TGAGTACTGAGGAGTTGAGGAAGTT 1545 Left TTAATCATTTCCCCTAATCCTTTTC
1546 Right TGAGGAAGTTCAATAAAGCTCACTT
1527 Left AACTTCAGCTTGGAGATAAAATCCT
1528 Right TCAATAAAGCTCACTTTGAAGGTCT 1547 Left TTAATCATTTCCCCTAATCCTTTTC
1548 Right TTGAGGAAGTTCAATAAAGCTCACT
1529 Left AACTTCAGCTTGGAGATAAAATCCT
1530 Right TTGAGGAAGTTCAATAAAGCTCACT 1549 Left AACTTCAGCTTGGAGATAAAATCCT
Seq. . 1550 Right CTGAGGAGTTGAGGAAGTTCAATAA
ID Pnmer Sequence 1531 Left AACTTCAGCTTGGAGATAAAATCCT 1551 Left GGGAAGGAACTATTTAAACTTCAGC
1532 Right TGAGGAAGTTCAATAAAGCTCACTT 1552 Right AGTATCCAAGTTATCCCATGTCTCA
1533 Left TTAATCATTTCCCCTAATCCTTTTC 1553 Left GGGAAGGAACTATTTAAACTTCAGC
1534 Right AGTATCCAAGTTATCCCATGTCTCA 1554 Right GTATCCAAGTTATCCCATGTCTCAG
1535 Left TTAATCATTTCCCCTAATCCTTTTC 1555 Left AACTTCAGCTTGGAGATAAAATCCT
1536 Right GTATCCAAGTTATCCCATGTCTCAG 1556 Right TTCAATAAAGCTCACTTTGAAGGTC
1537 Left TCCTAGGGATAAAATTAGGAAATGC 1557 Left AACTTCAGCTTGGAGATAAAATCCT
1538 Right AGTACTGAGGAGTTGAGGAAGTTCA 1558 Right GGAAGTTCAATAAAGCTCACTTTGA
1539 Left TCCTAGGGATAAAATTAGGAAATGC ¨1559 Left AACTTCAGCTTGGAGATAAAATCCT
1540 Right TGAGTACTGAGGAGTTGAGGAAGTT 1560 Right GTCTTTCCACATCAAGTATCCAAGT
1541 Left TTAATCATTTCCCCTAATCCTTTTC
ALK Exon25 2kb Seq. Pn .
mer Sequence ID 1575 Left AAATGAGGACAGTAATAGCACCTTG
1561 Left GGACAGTAATAGCACCTTGTGTCTT 1576 Right TGCATTGCAATATAGAAAACACAGT
1562 Right GTGCTGAGGACATAAATAGGTCAGT
1577 Left AAAATCATGGACAAAAGAACCATAA
1563 Left AAAATCATGGACAAAAGAACCATAA 1578 Right GGTGTCTGGATCAGTCTCACTTATT
1564 Right TTAAAGCTCCATATAACGATTGCTC
1579 Left AAAATCATGGACAAAAGAACCATAA
1565 Left GGACAGTAATAGCACCTTGTGTCTT 1580 Right GGAACTAGAGGCTAGGAAGAGAAGA
1566 Right AGTCTCTCTCTCCCAAGGATATTGT
__________________________________________ Seq. .
S
1567 Left AAAATCATGGACAAAAGAACCATAA ID Primer equen ce 1568 Right AGTCTCACTTATTCCCCAAAGAGTT 1581 Left AAATGAGGACAGTAATAGCACCTTG
1582 Right GTGCTGAGGACATAAATAGGTCAGT
1569 Left ATCACTTTTTAAAACAACCATTCCA
1570 Right TGGTCACTAATTAAGGTTTCCCATA 1583 Left AACTTCAGCTTGGAGATAAAATCCT
1584 Right TGCATTGCAATATAGAAAACACAGT
1571 Left TCACTTTTTAAAACAACCATTCCAT
1572 Right TGGTCACTAATTAAGGTTTCCCATA 1585 Left AAATGAGGACAGTAATAGCACCTTG
1586 Right AGTCTCTCTCTCCCAAGGATATTGT
1573 Left TGTAGCTTAGCAAGGGCTTTAGATA
1574 Right TTAAAGCTCCATATAACGATTGCTC 1587 Left GTCACTCTCCCAACTCTTGATGTAT

1588 Right TGGTCACTAATTAAGGTTTCCCATA
1595 Left TGTAGCTTAGCAAGGGCTTTAGATA
1589 Left AACTTCAGCTTGGAGATAAAATCCT 1596 Right GGAACTAGAGGCTAGGAAGAGAAGA
1590 Right GTGCTGAGGACATAAATAGGTCAGT
1597 Left GGACAGTAATAGCACCTTGTGTCTT
1591 Left TGTAGCTTAGCAAGGGCTTTAGATA 1598 Right TCAGTGACACAAATGAAGAATTGAT
1592 Right AGTCTCACTTATTCCCCAAAGAGTT
1599 Left TTAATCATTTCCCCTAATCCTTTTC
1593 Left TGTAGCTTAGCAAGGGCTTTAGATA 1600 Right TGCATTGCAATATAGAAAACACAGT
1594 Right GGTGTCTGGATCAGTCTCACTTATT
ALK Exon24-25 5kb Seq. . 1622 Right AGCCTGAAAAGGAACTTAGTGAAAT
ID Primer Sequence 1601 Left ATCTCCTTTTGAATGAAAGAGACCT
1602 Right TGGTCACTAATTAAGGTTTCCCATA
Seq. . 1623 Left TGCACAATAAATTAAAAGGGAAAGA

Primer Sequence ID 1624 Right TGGTCACTAATTAAGGTTTCCCATA
1603 Left ATCTCCTTTTGAATGAAAGAGACCT
1604 Right AGCCTGAAAAGGAACTTAGTGAAAT 1625 Left TGTTTATAAATTGGGGGTATTCAAA
1605 Left ACGTCAGGGATTTAGGAATTAAAAG 1626 Right TGGTCACTAATTAAGGTTTCCCATA
1606 Right TGGTCACTAATTAAGGTTTCCCATA
1627 Left TTGTTTATAAATTGGGGGTATTCAA
1607 Left ATGTGAATCATACTCCTCCAGGTAA 1628 Right TGGTCACTAATTAAGGTTTCCCATA
1608 Right TGGTCACTAATTAAGGTTTCCCATA
1629 Left TGAATCATACTCCTCCAGGTAAATC
1609 Left ATCTCCTTTTGAATGAAAGAGACCT 1630 Right TGGTCACTAATTAAGGTTTCCCATA
1610 Right GTATCCAAGTTATCCCATGTCTCAG
1631 Left AGGTATGTGAATCATACTCCTCCAG
1611 Left ATCTCCTTTTGAATGAAAGAGACCT 1632 Right AGCCTGAAAAGGAACTTAGTGAAAT
1612 Right TAGCCTGAAAAGGAACTTAGTGAAA
1633 Left ACGTCAGGGATTTAGGAATTAAAAG
1613 Left ACGTCAGGGATTTAGGAATTAAAAG 1634 Right TAGCCTGAAAAGGAACTTAGTGAAA
1614 Right AGCCTGAAAAGGAACTTAGTGAAAT
1635 Left AAGAGTCACCAGCTTAAACAAACAC
1615 Left ATCTCCTTTTGAATGAAAGAGACCT 1636 Right TAGCCTGAAAAGGAACTTAGTGAAA
1616 Right CATAGCCTGAAAAGGAACTTAGTGA
1637 Left ATGTGAATCATACTCCTCCAGGTAA
1617 Left AGGTATGTGAATCATACTCCTCCAG 1638 Right TAGCCTGAAAAGGAACTTAGTGAAA
1618 Right TGGTCACTAATTAAGGTTTCCCATA
1639 Left TGCACAATAAATTAAAAGGGAAAGA
1619 Left AAGAGTCACCAGCTTAAACAAACAC 1640 Right AGCCTGAAAAGGAACTTAGTGAAAT
1620 Right AGCCTGAAAAGGAACTTAGTGAAAT
1621 Left ATGTGAATCATACTCCTCCAGGTAA
Table 8. EGFR Capture Primer List for NGS Panel EGFR Exon18 100-200 bases Seq. . 1644 Right TTCTTGACGAGGTCCATGTG
ID Primer Sequence ¨
1641 Left TGCCAAAGAAGTAGAATGAG 1645 Left TGCCAAAGAAGTAGAATGAG
1642 Right AAAGCATCTTCACCCACAGC 1646 Right GTCAGAAATGCAGGAAAGCA
1643 Left TGCCAAAGAAGTAGAATGAG 1647 Left TGCCAAAGAAGTAGAATGAG

1648 Right AGTCAGAAATGCAGGAAAGCA 1664 Right AAAGCATCTTCACCCACAGC
1649 Left TGCCAAAGAAGTAGAATGAG 1665 Left TGCCAAAGAAGTAGAATGAG
1650 Right CAGTCAGAAATGCAGGAAAGC 1666 Right GGACAGTCAGAAATGCAGGAA
1651 Left TGCCAAAGAAGTAGAATGAG 1667 Left GCCAAAGAAGTAGAATGAGA
1652 Right ATTCTTGACGAGGTCCATGTG 1668 Right TTCTTGACGAGGTCCATGTG
1653 Left TGCCAAAGAAGTAGAATGAG 1669 Left TGCCAAAGAAGTAGAATGAG
1654 Right CATTCTTGACGAGGTCCATGT 1670 Right TGCAGGAAAGCATCTTCACC
Seq. .
Primer Sequence ID 1671 Left TGCCAAAGAAGTAGAATGAG
1655 Left TGCCAAAGAAGTAGAATGAG 1672 Right TCAGAAATGCAGGAAAGCATC
1656 Right GGAAAGCATCTTCACCCACA
1673 Left TGCCAAAGAAGTAGAATGAG
1657 Left TGCCAAAGAAGTAGAATGAG 1674 Right GTCATTCTTGACGAGGTCCA
1658 Right CAGCAGTGTGGTCATTCTTGA
1675 Left TGCCAAAGAAGTAGAATGAG
1659 Left TGCCAAAGAAGTAGAATGAG 1676 Right TGGTCATTCTTGACGAGGTC
1660 Right AGGACAGTCAGAAATGCAGGA
1677 Left TGCCAAAGAAGTAGAATGAG
1661 Left TGCCAAAGAAGTAGAATGAG 1678 Right AGCATCTTCACCCACAGCA
1662 Right GGACAGTCAGAAATGCAGGA
1679 Left TGCCAAAGAAGTAGAATGAG
1663 Left GCCAAAGAAGTAGAATGAGA 1680 Right GCATCTTCACCCACAGCAG
EGFR Exon18 200-400 bases Seq. . 1697 Left TGCCAAAGAAGTAGAATGAG
Primer Sequence ID 1698 Right AGCAAAACCAGTGGAACCAA
1681 Left TGCCAAAGAAGTAGAATGAG
1682 Right CCAGCACTGTGTGTCCAACT 1699 Left TGCCAAAGAAGTAGAATGAG
1700 Right AAACCAGTGGAACCAAGGAA
1683 Left TGCCAAAGAAGTAGAATGAG
1684 Right TCCCTCCACTGAGGACAAAG Seq. .
ID Pnmer Sequence 1685 Left TGCCAAAGAAGTAGAATGAG 1701 Left TGCCAAAGAAGTAGAATGAG
1686 Right CTTTCCCTCCACTGAGGACA 1702 Right AAAACCAGTGGAACCAAGGA
1687 Left TGCCAAAGAAGTAGAATGAG 1703 Left TGCCAAAGAAGTAGAATGAG
1688 Right CCAACTTTCCCTCCACTGAG 1704 Right GTGTCCAACTTTCCCTCCAC
1689 Left TGCCAAAGAAGTAGAATGAG 1705 Left TGCCAAAGAAGTAGAATGAG
1690 Right CAAAACCAGTGGAACCAAGG 1706 Right GGCCCAGAGCCATAGAAACT
1691 Left TGCCAAAGAAGTAGAATGAG 1707 Left TGCCAAAGAAGTAGAATGAG
1692 Right TGTCCAACTTTCCCTCCACT 1708 Right TTCCCTCCACTGAGGACAAA
1693 Left TGCCAAAGAAGTAGAATGAG 1709 Left TGCCAAAGAAGTAGAATGAG
1694 Right GTCCAACTTTCCCTCCACTG 1710 Right TCCAACTTTCCCTCCACTGA
1695 Left TGCCAAAGAAGTAGAATGAG 1711 Left TGCCAAAGAAGTAGAATGAG
1696 Right GCAAAACCAGTGGAACCAAG 1712 Right CCCTCCACTGAGGACAAAGT

1713 Left TGCCAAAGAAGTAGAATGAG 717 Left GCCAAAGAAGTAGAATGAGA
1714 Right AACCAGCTGGGCAGTCTCT 718 Right CCAGCACTGTGTGTCCAACT
1715 Left TGCCAAAGAAGTAGAATGAG 719 Left GCCAAAGAAGTAGAATGAGA
1716 Right GAAACCCTGGCTGAGGGTAG 720 Right TCCCTCCACTGAGGACAAAG
EGFR Exon18 400-1000 bases Seq. .
Primer Sequence ID 1739 Left TGCCAAAGAAGTAGAATGAG
1721 Left TGCCAAAGAAGTAGAATGAG 1740 Right ACGCCATCGAGAGTAACACC
1722 Right CAGTGTGGAGTGGGGAAGTT
1741 Left TGCCAAAGAAGTAGAATGAG
1723 Left TGCCAAAGAAGTAGAATGAG 1742 Right AGGAGCATGCCAAAATGAAG
1724 Right ACTCCCCTATGCTGGAGGTT
1743 Left TGCCAAAGAAGTAGAATGAG
1744 Right TGTTGAAGGAAGCCCTTTTG
Seq. . 1745 Left TGCCAAAGAAGTAGAATGAG
Primer Sequence ID 1746 Right CCAATGGGGTAAGTGGACAG
1725 Left TGCCAAAGAAGTAGAATGAG
1726 Right TGGGAAAGAAAGCAAGGAGA 1747 Left TGCCAAAGAAGTAGAATGAG
1748 Right TTGCCTTCTTCCTCGATCAT
1727 Left TGCCAAAGAAGTAGAATGAG
1728 Right TCTGGGAAAGAAAGCAAGGA 1749 Left TGCCAAAGAAGTAGAATGAG
1750 Right CATCGAACAGAAAGGCCACT
__________________________________________ 1751 Left TGCCAAAGAAGTAGAATGAG
1729 Left TGCCAAAGAAGTAGAATGAG 1752 Right GGTGGCAGGAGAGAGAGCTA
1730 Right ACCAATGGGGTAAGTGGACA
¨1753 Left TGCCAAAGAAGTAGAATGAG
1731 Left TGCCAAAGAAGTAGAATGAG 1754 Right ATGGGACCAATGGGGTAAGT
1732 Right CCTCGATCATGTGACACTGG
7 755 Left TGCCAAAGAAGTAGAATGAG
1733 Left TGCCAAAGAAGTAGAATGAG 1756 Right TGGAGGTTGTCATCGAACAG
1734 Right AAAATGGCAAACAGGTGCTC
¨1757 Left TGCCAAAGAAGTAGAATGAG
1735 Left TGCCAAAGAAGTAGAATGAG 1758 Right CTGGAGGTTGTCATCGAACA
1736 Right AACTGGCCAGAGCTGATGTT
¨1759 Left TGCCAAAGAAGTAGAATGAG
1737 Left TGCCAAAGAAGTAGAATGAG 1760 Right TATGCTGGAGGTTGTCATCG
1738 Right AAACTGGCCAGAGCTGATGT
EGFR Exon19 100-200 bases Seq. .
Pnmer Sequence ID 1767 Left CTTGTTCCTCCACCTCATTCC
1761 Left CTTCCTTGTTCCTCCACCTCAT 1768 Right ACCCAGGACTGGCACTCAC
1762 Right ACCCAGGACTGGCACTCAC
1769 Left CCTTGTTCCTCCACCTCATTC
1763 Left CTTCCTTGTTCCTCCACCTCAT 1770 Right ACCCAGGACTGGCACTCAC
1764 Right CCCAGGACTGGCACTCAC
1771 Left TCCTTGTTCCTCCACCTCATT
1765 Left CTTCCTTGTTCCTCCACCTCAT 1772 Right ACCCAGGACTGGCACTCAC
1766 Right ACCCAGGACTGGCACTCA

1773 Left TTCCTTGTTCCTCCACCTCAT
1774 Right ACCCAGGACTGGCACTCAC 1787 Left TCCTTGTTCCTCCACCTCATT
1788 Right CCCAGGACTGGCACTCAC
1775 Left CTTCCTTGTTCCTCCACCTCATT
1776 Right ACCCAGGACTGGCACTCAC 1789 Left TCCTTGTTCCTCCACCTCATT
1790 Right ACCCAGGACTGGCACTCA
1777 Left CAACCTCACCCTTCCTTGTTC
1778 Right ACCCAGGACTGGCACTCAC 1791 Left TTCCTTGTTCCTCCACCTCAT
Seq. . 1792 Right CCCAGGACTGGCACTCAC
ID Pnmer Sequence ¨
1779 Left CTTGTTCCTCCACCTCATTCC 1793 Left TTCCTTGTTCCTCCACCTCAT
1780 Right CCCAGGACTGGCACTCAC 1794 Right ACCCAGGACTGGCACTCA
1781 Left CCTTGTTCCTCCACCTCATTC 1795 Left CTTCCTTGTTCCTCCACCTCATT
1782 Right CCCAGGACTGGCACTCAC 1796 Right CCCAGGACTGGCACTCAC
1783 Left CCTTGTTCCTCCACCTCATTC 1797 Left CTTCCTTGTTCCTCCACCTCATT
1784 Right ACCCAGGACTGGCACTCA 1798 Right ACCCAGGACTGGCACTCA
1785 Left CTTGTTCCTCCACCTCATTCC 1799 Left CAACCTCACCCTTCCTTGTTC
1786 Right ACCCAGGACTGGCACTCA 1800 Right CCCAGGACTGGCACTCAC
EGFR Exon19 200-400 bases Seq.
Primer Sequence ID Seq.
Primer Sequence 1801 Left AAGATCATTCTACAAGATGTCAGTGG ID
1802 Right AACTGCACATTCAGAGATTCTTTCT 1821 Left AGATCATTCTACAAGATGTCAGTGC
1822 Right TAACTGCACATTCAGAGATTCTTTC
1803 Left AGATCATTCTACAAGATGTCAGTGC
1804 Right AACTGCACATTCAGAGATTCTTTCT 1823 Left AAGATCATTCTACAAGATGTCAGTGC
1824 Right TAACTGCACATTCAGAGATTCTTTCT
1805 Left TCCAAGATCATTCTACAAGATGTCA
1806 Right ACATTCAGAGATTCTTTCTGCATCA 1825 Left AGATCATTCTACAAGATGTCAGTGC
1826 Right TAACTGCACATTCAGAGATTCTTTCT
1807 Left TCCAAGATCATTCTACAAGATGTCA
1808 Right ATTCAGAGATTCTTTCTGCATCATA 1827 Left TCCAAGATCATTCTACAAGATGTCA
1828 Right TTCAGAGATTCTTTCTGCATCATAATA
1809 Left TCCAAGATCATTCTACAAGATGTCA
1810 Right CATTCAGAGATTCTTTCTGCATCA 1829 Left TCCAAGATCATTCTACAAGATGTCA
1830 Right ACTGCACATTCAGAGATTCTTTCT
1811 Left CAAGATCATTCTACAAGATGTCAGTG
1812 Right AACTGCACATTCAGAGATTCTTTCT 1831 Left TCCAAGATCATTCTACAAGATGTCA
1832 Right GCACATTCAGAGATTCTTTCTGC
1813 Left TCCAAGATCATTCTACAAGATGTCA
1814 Right CATTCAGAGATTCTTTCTGCATCATA 1833 Left TCCAAGATCATTCTACAAGATGTCA
1834 Right TCAGAGATTCTTTCTGCATCATAATA
1815 Left AAGATCATTCTACAAGATGTCAGTGG
1816 Right CATTCAGAGATTCTTTCTGCATCAT 1835 Left CCAAGATCATTCTACAAGATGTCAGT
1836 Right ACATTCAGAGATTCTTTCTGCATCA
1817 Left AAGATCATTCTACAAGATGTCAGTGG
1818 Right TAACTGCACATTCAGAGATTCTTTC 1837 Left CCAAGATCATTCTACAAGATGTCAGT
1838 Right ATTCAGAGATTCTTTCTGCATCATA
1819 Left AGATCATTCTACAAGATGTCAGTGC
1820 Right CATTCAGAGATTCTTTCTGCATCAT 1839 Left AAGATCATTCTACAAGATGTCAGTGC

1840 Right ACATTCAGAGATTCTTTCTGCATCA
EGFR Exon19 400-1000 bases Seq.
Primer Sequence ID 1861 Left CTCGTTCAGAGAGTATTTCACACAA

1841 Left AATACCAATCCATGAAAAAGCATTA 1862 Right AACATGTCACCAACTGGGTATAACT
1842 Right AACATGTCACCAACTGGGTATAACT
1863 Left TCCAAGATCATTCTACAAGATGTCA
1843 Left CCTATTCCTTTATAACCCCTTTCAA ¨1864 Right ACTGAACAGCTACCTTTCAACAAAC
1844 Right AACATGTCACCAACTGGGTATAACT
1865 Left CCTATTCCTTTATAACCCCTTTCAA
1845 Left TCCAAGATCATTCTACAAGATGTCA 1866 Right AACTGCACATTCAGAGATTCTTTCT
1846 Right AACATGTCACCAACTGGGTATAACT
1867 Left TCCAAGATCATTCTACAAGATGTCA
1847 Left TTTCAAGCTCGTTCAGAGAGTATTT 1868 Right AACTGCACATTCAGAGATTCTTTCT
1848 Right AACATGTCACCAACTGGGTATAACT
1869 Left ACTCTTGGAATGAACAAAATACCAA
1849 Left TTCAGAGAGTATTTCACACAATCCA 1870 Right AACATGTCACCAACTGGGTATAACT
1850 Right AACATGTCACCAACTGGGTATAACT ____ ¨
Seq. 1871 Left CCTATTCCTTTATAACCCCTTTCAA

Primer Sequence ID 1872 Right GGGTATAACTGCACATTCAGAGATT
1851 Left GTGTCTCACTTTCCAAGATCATTCT _____ 1852 Right AACATGTCACCAACTGGGTATAACT 1873 Left TCCAAGATCATTCTACAAGATGTCA
1874 Right GGGTATAACTGCACATTCAGAGATT
1853 Left AGTGTCTCACTTTCCAAGATCATTC _____ 1854 Right AACATGTCACCAACTGGGTATAACT 1875 Left AGTGTCTCACTTTCCAAGATCATTC
1876 Right ACTGAACAGCTACCTTTCAACAAAC
1855 Left CCATGAAAAAGCATTATTGAAGTCT _____ 1856 Right AACATGTCACCAACTGGGTATAACT 1877 Left GTGTCTCACTTTCCAAGATCATTCT
1878 Right ACTGAACAGCTACCTTTCAACAAAC
1857 Left TATTCCTTTATAACCCCTTTCAAGC _____ 1858 Right AACATGTCACCAACTGGGTATAACT 1879 Left AGTGTCTCACTTTCCAAGATCATTC
1880 Right AACTGCACATTCAGAGATTCTTTCT
1859 Left ATGGAAATACTCTTGGAATGAACAA
1860 Right AACATGTCACCAACTGGGTATAACT
EGFR Exon20 100-200 bases Seq. 1891 Left AGGTGACCCTTGTCTCTGTGTT
Primer Sequence ID 1892 Right CCTGTGCCAGGGACCTTAC
1881 Left GTGACCCTTGTCTCTGTGTTCTT
1882 Right CCTGTGCCAGGGACCTTAC 1893 Left GACCCTTGTCTCTGTGTTCTTGT
1894 Right CCTGTGCCAGGGACCTTA
1883 Left ACCCTTGTCTCTGTGTTCTTGTC
1884 Right CCTGTGCCAGGGACCTTAC 1895 Left GTGACCCTTGTCTCTGTGTTCTT
¨1896 Right CCTGTGCCAGGGACCTTA
1885 Left GACCCTTGTCTCTGTGTTCTTGT
1886 Right CCTGTGCCAGGGACCTTAC 1897 Left ACCCTTGTCTCTGTGTTCTTGTC
1898 Right CCTGTGCCAGGGACCTTA
1887 Left GACCCTTGTCTCTGTGTTCTTGTC _____ ¨
1888 Right CCTGTGCCAGGGACCTTAC 1899 Left GACCCTTGTCTCTGTGTTCTTGTC
1900 Right CCTGTGCCAGGGACCTTA
1889 Left GTGACCCTTGTCTCTGTGTTCTTGT
1890 Right CCTGTGCCAGGGACCTTAC Seq.
ID Primer Sequence 1901 Left GTGACCCTTGTCTCTGTGTTCTTG 911 Left CTGAGGTGACCCTTGTCTCTGT
1902 Right CCTGTGCCAGGGACCTTAC 912 Right CCTGTGCCAGGGACCTTAC
1903 Left TGACCCTTGTCTCTGTGTTCTTGT 913 Left AGGTGACCCTTGTCTCTGTGTTCTT
1904 Right CCTGTGCCAGGGACCTTAC 914 Right CCTGTGCCAGGGACCTTAC
1905 Left GAGGTGACCCTTGTCTCTGTGT 915 Left GGTGACCCTTGTCTCTGTGTTCT
1906 Right CCTGTGCCAGGGACCTTAC 916 Right CCTGTGCCAGGGACCTTAC
1907 Left TGACCCTTGTCTCTGTGTTCTTG 917 Left AGGTGACCCTTGTCTCTGTGTTC
1908 Right CCTGTGCCAGGGACCTTAC 918 Right CCTGTGCCAGGGACCTTAC
1909 Left AGGTGACCCTTGTCTCTGTGTTCT 919 Left GAGGTGACCCTTGTCTCTGTGTT
1910 Right CCTGTGCCAGGGACCTTAC 920 Right CCTGTGCCAGGGACCTTAC
EGFR Exon20 200-400 bases Seq. Primer Sequence ID 1941 Left TACATTTGTCCTTCCAAATGAGC
1921 Left AAGCTCTGTAGAGAAGGCGTACAT 1942 Right AAATATACAGCTTGCAAGGACTCTG
1922 Right AAATATACAGCTTGCAAGGACTCTG
Seq. . 1943 Left CAAGCTCTGTAGAGAAGGCGTACAT

Primer Sequence ID 1944 Right AAATATACAGCTTGCAAGGACTCTG
1923 Left AGCTCTGTAGAGAAGGCGTACATT
1924 Right AAATATACAGCTTGCAAGGACTCTG 1945 Left AGCTCTGTAGAGAAGGCGTACATT
1925 Left TCTGTAGAGAAGGCGTACATTTGTC 1946 Right GGAAATATACAGCTTGCAAGGACTC
1926 Right AAATATACAGCTTGCAAGGACTCTG _____ 1947 Left AAGCTCTGTAGAGAAGGCGTACAT
1927 Left GTAGAGAAGGCGTACATTTGTCCT 1948 Right GGAAATATACAGCTTGCAAGGACTC
1928 Right AAATATACAGCTTGCAAGGACTCTG _____ 1949 Left GTCAAGCTCTGTAGAGAAGGCGTA
1929 Left AAGCTCTGTAGAGAAGGCGTACATT 1950 Right AAATATACAGCTTGCAAGGACTCTG
1930 Right AAATATACAGCTTGCAAGGACTCTG _____ 1951 Left TCTGTAGAGAAGGCGTACATTTGTC
1931 Left GCTCTGTAGAGAAGGCGTACATTT 1952 Right GGAAATATACAGCTTGCAAGGACTC
1932 Right AAATATACAGCTTGCAAGGACTCTG _____ 1953 Left TGTAGAGAAGGCGTACATTTGTCCT
1933 Left CTCTGTAGAGAAGGCGTACATTTG 1954 Right AAATATACAGCTTGCAAGGACTCTG
1934 Right AAATATACAGCTTGCAAGGACTCTG _____ 1955 Left GTAGAGAAGGCGTACATTTGTCCT
1935 Left CTGTAGAGAAGGCGTACATTTGTC 1956 Right GGAAATATACAGCTTGCAAGGACTC
1936 Right AAATATACAGCTTGCAAGGACTCTG _____ 1957 Left AAGCTCTGTAGAGAAGGCGTACATT
1937 Left TTCTGTCAAGCTCTGTAGAGAAGG 1958 Right GGAAATATACAGCTTGCAAGGACTC
1938 Right AAATATACAGCTTGCAAGGACTCTG _____ 1959 Left AGCTCTGTAGAGAAGGCGTACATT
1939 Left AAGCTCTGTAGAGAAGGCGTACA 1960 Right ATGGAAATATACAGCTTGCAAGGAC
1940 Right AAATATACAGCTTGCAAGGACTCTG
EGFR Exon20 400-1000 bases Seq. Pn .
mer Sequence ID 1963 Left GTTTCTACCAACTTCTGTCAAGCTC

1961 Left TTTCTACCAACTTCTGTCAAGCTCT 1964 Right ATCTAGAAGAAGCAAACGAAGATGA
1962 Right ATCTAGAAGAAGCAAACGAAGATGA

1965 Left GTTTCTACCAACTTCTGTCAAGCTC 1983 Left AATCTCTGAATGTGCAGTTATACCC
1966 Right GATCTAGAAGAAGCAAACGAAGATG 1984 Right AAATATACAGCTTGCAAGGACTCTG
1967 Left GTTTCTACCAACTTCTGTCAAGCTC 1985 Left TTTCTACCAACTTCTGTCAAGCTCT
1968 Right CTATGACAGAGAGAGAAGGAAGACC 1986 Right GTTATAAAGTCCGTGTGGATCATTT
1969 Left CTGTGTTTCTACCAACTTCTGTCAA 1987 Left CTGTGTTTCTACCAACTTCTGTCAA
1970 Right GATCTAGAAGAAGCAAACGAAGATG 1988 Right AAATATACAGCTTGCAAGGACTCTG
1971 Left TGTGTTTCTACCAACTTCTGTCAAG 1989 Left TTTCTACCAACTTCTGTCAAGCTCT
1972 Right ATCTAGAAGAAGCAAACGAAGATGA 1990 Right TTATAAAGTCCGTGTGGATCATTTC
1973 Left TGTGTTTCTACCAACTTCTGTCAAG 1991 Left GAAATTGTGTTTGTTGAAAGGTAGC
1974 Right GATCTAGAAGAAGCAAACGAAGATG 1992 Right AAATATACAGCTTGCAAGGACTCTG
1975 Left CTGTGTTTCTACCAACTTCTGTCAA 1993 Left TTTCTACCAACTTCTGTCAAGCTCT
1976 Right ATCTAGAAGAAGCAAACGAAGATGA 1994 Right CTGTTATAAAGTCCGTGTGGATCAT
S eq. .
Primer S equence ID 1995 Left GTTTCTACCAACTTCTGTCAAGCTC

1977 Left AGAAAGAATCTCTGAATGTGCAGTT 1996 Right GTTATAAAGTCCGTGTGGATCATTT
1978 Right AAATATACAGCTTGCAAGGACTCTG
1997 Left TTTCTACCAACTTCTGTCAAGCTCT
1979 Left GAAATTGTGTTTGTTGAAAGGTAGC 1998 Right TCTAGAAGAAGCAAACGAAGATGAG
1980 Right ATCTAGAAGAAGCAAACGAAGATGA
1999 Left TTTCTACCAACTTCTGTCAAGCTCT
1981 Left GAAATTGTGTTTGTTGAAAGGTAGC 2000 Right CTCCACGAATCACACTGATTATTTA
1982 Right GATCTAGAAGAAGCAAACGAAGATG
EGFR Exon21 100-200 bases S eq. . 2016 Right ACACAGCAAAGCAGAAACTCAC
ID Pnmer Sequence 2001 Left ACGTCTTCCTTCTCTCTCTGTCATA 2017 Left CAGTTAACGTCTTCCTTCTCTCTCT
2002 Right ACACAGCAAAGCAGAAACTCAC 2018 Right ACACAGCAAAGCAGAAACTCAC
2003 Left TTAACGTCTTCCTTCTCTCTCTGTC 72019 Left AGTTAACGTCTTCCTTCTCTCTCTG
2004 Right ACACAGCAAAGCAGAAACTCAC 2020 Right ACACAGCAAAGCAGAAACTCAC
2005 Left TAACGTCTTCCTTCTCTCTCTGTCA Seq. .
S
2006 Right ACACAGCAAAGCAGAAACTCAC ID Primer equence 2021 Left ACGTCTTCCTTCTCTCTCTGTCATA
2007 Left ACGTCTTCCTTCTCTCTCTGTCATA 2022 Right CACACAGCAAAGCAGAAACTCA
2008 Right CACACAGCAAAGCAGAAACTCAC
2023 Left GTTAACGTCTTCCTTCTCTCTCTGT
2009 Left ACGTCTTCCTTCTCTCTCTGTCAT 2024 Right ACACAGCAAAGCAGAAACTCAC
2010 Right ACACAGCAAAGCAGAAACTCAC
2025 Left TTAACGTCTTCCTTCTCTCTCTGTC
2011 Left CCAGTTAACGTCTTCCTTCTCTCTC 2026 Right CACACAGCAAAGCAGAAACTCAC
2012 Right ACACAGCAAAGCAGAAACTCAC
2027 Left GTTAACGTCTTCCTTCTCTCTCTGTC
2013 Left ACGTCTTCCTTCTCTCTCTGTCATA 2028 Right ACACAGCAAAGCAGAAACTCAC
2014 Right CCACACAGCAAAGCAGAAACT
2029 Left AGTTAACGTCTTCCTTCTCTCTCTGT
2015 Left AACGTCTTCCTTCTCTCTCTGTCAT 2030 Right ACACAGCAAAGCAGAAACTCAC

2036 Right CACACAGCAAAGCAGAAACTCA
2031 Left TTAACGTCTTCCTTCTCTCTCTGTC
2032 Right CCACACAGCAAAGCAGAAACT 2037 Left TAACGTCTTCCTTCTCTCTCTGTCA

2038 Right CACACAGCAAAGCAGAAACTCAC
2033 Left CCAGTTAACGTCTTCCTTCTCTCT
2034 Right ACACAGCAAAGCAGAAACTCAC 2039 Left CGTCTTCCTTCTCTCTCTGTCATA
2040 Right ACACAGCAAAGCAGAAACTCAC
2035 Left TTAACGTCTTCCTTCTCTCTCTGTC
EGFR Exon21 200-400 bases S eq. .
Primer S equence ID 2061 Left ACGTCTTCCTTCTCTCTCTGTCATA

2041 Left ACGTCTTCCTTCTCTCTCTGTCATA 2062 Right AAGTGAACATTTAGGATGTGGAGAT
2042 Right TGTCTCTAAGGGGAGGGAGTTATAC
2063 Left ACGTCTTCCTTCTCTCTCTGTCATA
2043 Left ACGTCTTCCTTCTCTCTCTGTCATA 2064 Right GTCAAGAAACTAGTGCTGGGTAGAT
2044 Right GAAAGTGAACATTTAGGATGTGGAG
2065 Left TTAACGTCTTCCTTCTCTCTCTGTC
2045 Left ACGTCTTCCTTCTCTCTCTGTCATA 2066 Right GTGTCAAGAAACTAGTGCTGGGTAG
2046 Right AAAGTGAACATTTAGGATGTGGAGA
2067 Left TTAACGTCTTCCTTCTCTCTCTGTC
2047 Left ACGTCTTCCTTCTCTCTCTGTCATA 2068 Right AGTGAACATTTAGGATGTGGAGATG
2048 Right AGAAAGTGAACATTTAGGATGTGGA
S eq. . 2069 Left TTAACGTCTTCCTTCTCTCTCTGTC

Pnmer S equence ID 2070 Right AAGTGAACATTTAGGATGTGGAGAT
2049 Left TTAACGTCTTCCTTCTCTCTCTGTC _____ 2050 Right TGTCTCTAAGGGGAGGGAGTTATAC 2071 Left ACGTCTTCCTTCTCTCTCTGTCATA
2072 Right TCAAGAAACTAGTGCTGGGTAGATG
2051 Left ACGTCTTCCTTCTCTCTCTGTCATA _____ 2052 Right GTGTCAAGAAACTAGTGCTGGGTAG 2073 Left TTAACGTCTTCCTTCTCTCTCTGTC
2074 Right GTCAAGAAACTAGTGCTGGGTAGAT
2053 Left TTAACGTCTTCCTTCTCTCTCTGTC _____ 2054 Right AAAGTGAACATTTAGGATGTGGAGA 2075 Left ACGTCTTCCTTCTCTCTCTGTCATA
AA. G AGGGAAAGACATAGAAAGTGAA
2055 Left TTAACGTCTTCCTTCTCTCTCTGTC 2076 RightC
2056 Right GAAAGTGAACATTTAGGATGTGGAG
2077 Left ACGTCTTCCTTCTCTCTCTGTCATA
2057 Left TTAACGTCTTCCTTCTCTCTCTGTC 2078 Right TGTCAAGAAACTAGTGCTGGGTAG
2058 Right AGAAAGTGAACATTTAGGATGTGGA
2079 Left TAACGTCTTCCTTCTCTCTCTGTCA
2059 Left ACGTCTTCCTTCTCTCTCTGTCATA 2080 Right TGTCTCTAAGGGGAGGGAGTTATAC
2060 Right AGTGAACATTTAGGATGTGGAGATG
EGFR Exon21 400-1000 bases S eq. .
Primer S equence ID 2087 Left ACGTCTTCCTTCTCTCTCTGTCATA

2081 Left ACGTCTTCCTTCTCTCTCTGTCATA 2088 Right TGAGGTAATAAGTCAGCCATTTTTC
2082 Right CAAAGTAACAATCAACAGACACTGG
_________________________________________ 2089 Left ACGTCTTCCTTCTCTCTCTGTCATA

2083 Left ACGTCTTCCTTCTCTCTCTGTCATA 2090 Right CCACAAAGTAACAATCAACAGACAC
2084 Right AAAGATGAGATAACTTGGTGGAGTG ____ 2091 Left TTCTAGATCCTCTTTGCATGAAATC
2085 Left ACGTCTTCCTTCTCTCTCTGTCATA 2092 Right AAAGATGAGATAACTTGGTGGAGTG
2086 Right CAAAGATGAGATAACTTGGTGGAGT

2093 Left TTCTAGATCCTCTTTGCATGAAATC
2094 Right CAAAGATGAGATAACTTGGTGGAGT 2107 Left ACGTCTTCCTTCTCTCTCTGTCATA
2108 Right CCATTTCAAAGATGAGATAACTTGG
2095 Left ACGTCTTCCTTCTCTCTCTGTCATA
2096 Right GGTAATAAGTCAGCCATTTTTCCTT 2109 Left AGGCTTTACAAGCTTGAGATTCTTT
2110 Right CAAAGTAACAATCAACAGACACTGG
2097 Left ACGTCTTCCTTCTCTCTCTGTCATA
2098 Right AATTTCTTTATGCCTCCATTTCTTC 2111 Left TTCTAGATCCTCTTTGCATGAAATC
2112 Right AATTTCTTTATGCCTCCATTTCTTC
2099 Left ACGTCTTCCTTCTCTCTCTGTCATA
2100 Right GATGAGATAACTTGGTGGAGTGAAT 2113 Left ACGTCTTCCTTCTCTCTCTGTCATA
2114 Right AGATAACTTGGTGGAGTGAATTGAA
Seq. .
Pnmer Sequence ID 2115 Left TTCTAGATCCTCTTTGCATGAAATC

2101 Left ACGTCTTCCTTCTCTCTCTGTCATA 2116 Right GATGAGATAACTTGGTGGAGTGAAT
2102 Right GAATTTTCCAAGAACTTATTCCACA
2117 Left TTCTAGATCCTCTTTGCATGAAATC
2103 Left ACGTCTTCCTTCTCTCTCTGTCATA 2118 Right CCATTTCAAAGATGAGATAACTTGG
2104 Right TGTGGAATTTTCCAAGAACTTATTC
2119 Left AGGCTTTACAAGCTTGAGATTCTTT
2105 Left ACGTCTTCCTTCTCTCTCTGTCATA 2120 Right AAAGATGAGATAACTTGGTGGAGTG
2106 Right AATAAGTCAGCCATTTTTCCTTTTC
EGFR Exon22 100-200 bases Seq. . 2140 Right CCGTATCTCCCTTCCCTGATTAC
ID Primer Sequence 2121 Left CACACTGACGTGCCTCTCC Seq. .
2122 Right CGTATCTCCCTTCCCTGATTAC ID Primer Sequence 2141 Left CGAAGCCACACTGACGTG
2123 Left CCACACTGACGTGCCTCTC 2142 Right CCGTATCTCCCTTCCCTGATTA
2124 Right CGTATCTCCCTTCCCTGATTAC
2143 Left CATGCGAAGCCACACTGAC
2125 Left CACACTGACGTGCCTCTCC 2144 Right CGTATCTCCCTTCCCTGATTAC
2126 Right CCGTATCTCCCTTCCCTGATTAC
2145 Left CATGCGAAGCCACACTGA
2127 Left CCACACTGACGTGCCTCTC 2146 Right CGTATCTCCCTTCCCTGATTAC
2128 Right CCGTATCTCCCTTCCCTGATTAC
2147 Left CATGCGAAGCCACACTGAC
2129 Left CACACTGACGTGCCTCTCC 2148 Right CCGTATCTCCCTTCCCTGATTAC
2130 Right CCGTATCTCCCTTCCCTGATTA
2149 Left CACACTGACGTGCCTCTCC
2131 Left CCACACTGACGTGCCTCTC 2150 Right TATCTCCCCTCCCCGTATCT
2132 Right CCGTATCTCCCTTCCCTGATTA
2151 Left CATGCGAAGCCACACTGA
2133 Left CGAAGCCACACTGACGTG 2152 Right CCGTATCTCCCTTCCCTGATTAC
2134 Right CGTATCTCCCTTCCCTGATTAC
2153 Left CACACTGACGTGCCTCTCC
2135 Left CGAAGCCACACTGACGTG 2154 Right CCGTATCTCCCTTCCCTGAT
2136 Right CCGTATCTCCCTTCCCTGATTAC
2155 Left CCACACTGACGTGCCTCTC
2137 Left AC CATGCGAAGCCACACT 2156 Right CCGTATCTCCCTTCCCTGAT
2138 Right CGTATCTCCCTTCCCTGATTAC
2157 Left ACCATGCGAAGCCACACT
2139 Left AC CATGCGAAGCCACACT 2158 Right CCGTATCTCCCTTCCCTGATTA

2160 Right TATCTCCCCTCCCCGTATCTC
2159 Left CACACTGACGTGCCTCTCC
EGFR Exon22 200-400 bases S eq. . 2180 Right CCGTATCTCCCTTCCCTGATTAC

ID Pnmer Sequence 2161 Left GTATTTTGAAACTCAAGATCGCATT
2162 Right CGTATCTCCCTTCCCTGATTAC
2181 Left CCATGAGTACGTATTTTGAAACTCA
2163 Left CCTCCATGAGTACGTATTTTGAAAC 2182 Right CATGGCAAACTCTTGCTATCC
2164 Right CGTATCTCCCTTCCCTGATTAC
2183 Left TATTTTGAAACTCAAGATCGCATTC
2165 Left GTATTTTGAAACTCAAGATCGCATT 2184 Right CGTATCTCCCTTCCCTGATTAC
2166 Right CCGTATCTCCCTTCCCTGATTAC
2185 Left GTATTTTGAAACTCAAGATCGCATT
2167 Left CCTCCATGAGTACGTATTTTGAAAC 2186 Right CTTATCTCCCCTCCCCGTATCT
2168 Right CCGTATCTCCCTTCCCTGATTAC
2187 Left GTATTTTGAAACTCAAGATCGCATT
2169 Left GTATTTTGAAACTCAAGATCGCATT 188 Right ACATATCCCCATGGCAAACTCT
2170 Right CATGGCAAACTCTTGCTATCC
___________________________________________ 189 Left CTTTTCCTCCATGAGTACGTATTTT

2190 Right CGTATCTCCCTTCCCTGATTAC
S eq. . 2191 Left TATTTTGAAACTCAAGATCGCATTC
Primer Sequence ID 2192 Right CCGTATCTCCCTTCCCTGATTAC

2171 Left GTATTTTGAAACTCAAGATCGCATT ______ 2172 Right ATATCCCCATGGCAAACTCTT 2193 Left GTATTTTGAAACTCAAGATCGCATT
2194 Right ACATATCCCCATGGCAAACTCTT
2173 Left CCATGAGTACGTATTTTGAAACTCA ______ 2174 Right CGTATCTCCCTTCCCTGATTAC 2195 Left CCTCCATGAGTACGTATTTTGAAAC
2196 Right CTTATCTCCCCTCCCCGTATCT
2175 Left GTATTTTGAAACTCAAGATCGCATT ______ 2176 Right CCGTATCTCCCTTCCCTGATTA 2197 Left CCATGAGTACGTATTTTGAAACTCA
2198 Right CCGTATCTCCCTTCCCTGATTA
2177 Left CCTCCATGAGTACGTATTTTGAAAC ______ 2178 Right CCGTATCTCCCTTCCCTGATTA 2199 Left GTATTTTGAAACTCAAGATCGCATT
2200 Right ACATATCCCCATGGCAAACTC
2179 Left CCATGAGTACGTATTTTGAAACTCA
EGFR Exon22 400-1000 bases S eq. . 2209 Left GTATTTTGAAACTCAAGATCGCATT
Pnmer Sequence ID 2210 Right ATCCAAAATAAAGGAATGTGTGTGT
2201 Left GTATTTTGAAACTCAAGATCGCATT
2202 Right TTGAATCCAAAATAAAGGAATGTGT 2211 Left GTATTTTGAAACTCAAGATCGCATT
2212 Right GCTTACCTTGTTATCAAGTCCTGAA
2203 Left GTATTTTGAAACTCAAGATCGCATT ______ 2204 Right CACACTGAGCACTCAATAAAGAGAA 2213 Left TGGTCTATTGAAAGAGCTTATCCAG
2214 Right TTGAATCCAAAATAAAGGAATGTGT
2205 Left GTATTTTGAAACTCAAGATCGCATT
2206 Right TTCTCCACTACAAATCACCACAGTA 2215 Left CCTCCATGAGTACGTATTTTGAAAC
2216 Right TTGAATCCAAAATAAAGGAATGTGT
2207 Left GTATTTTGAAACTCAAGATCGCATT
2208 Right ATTCTTCAAAGGTAGCTGATTGATG 2217 Left CCTCCATGAGTACGTATTTTGAAAC
2218 Right CACACTGAGCACTCAATAAAGAGAA

2229 Left GTATTTTGAAACTCAAGATCGCATT
2219 Left CCTCCATGAGTACGTATTTTGAAAC 2230 Right TATTCCTTCTCCACTACAAATCACC
2220 Right TTCTCCACTACAAATCACCACAGTA
2231 Left GTATTTTGAAACTCAAGATCGCATT
S eq. . 2232 Right CCACTACAAATCACCACAGTATTCA
ID Pnmer Sequence 2221 Left TGGTCTATTGAAAGAGCTTATCCAG 2233 Left GTATTTTGAAACTCAAGATCGCATT
2222 Right ATCCAAAATAAAGGAATGTGTGTGT 2234 Right CTTGATTGAATCCAAAATAAAGGAA
2223 Left CCTCCATGAGTACGTATTTTGAAAC 2235 Left GTATTTTGAAACTCAAGATCGCATT
2224 Right ATCCAAAATAAAGGAATGTGTGTGT 2236 Right TAAGAACAGAGACATCAGACCACAC
2225 Left CCTCCATGAGTACGTATTTTGAAAC 2237 Left CCTCCATGAGTACGTATTTTGAAAC
2226 Right GCTTACCTTGTTATCAAGTCCTGAA 2238 Right CAAAGGTAGCTGATTGATGAGAGTT
2227 Left GTATTTTGAAACTCAAGATCGCATT 2239 Left GTATTTTGAAACTCAAGATCGCATT
2228 Right CAAAGGTAGCTGATTGATGAGAGTT 2240 Right AAATTCTTCAAAGGTAGCTGATTGA
EGFR Exon18-19 2kb Seq. .
Primer S equence ID 2261 Left AATCTCCAAAATATATGCCAAAGAA

2241 Left ATGCCAAAGAAGTAGAATGAGAAAA 2262 Right GGGTATAACTGCACATTCAGAGATT
2242 Right AACATGTCACCAACTGGGTATAACT ____ S eq. . 2263 Left AAATCTCCAAAATATATGCCAAAGA

Pnmer S equence ID 2264 Right GGGTATAACTGCACATTCAGAGATT
2243 Left ATGCCAAAGAAGTAGAATGAGAAAA ______ 2244 Right GGGTATAACTGCACATTCAGAGATT 2265 Left ATCTCCAAAATATATGCCAAAGAAG
2266 Right AACATGTCACCAACTGGGTATAACT
2245 Left TCTCCAAAATATATGCCAAAGAAGT ______ 2246 Right AACATGTCACCAACTGGGTATAACT 2267 Left AAAAATCTCCAAAATATATGCCAAAG
2268 Right AACATGTCACCAACTGGGTATAACT
2247 Left TGCCAAAGAAGTAGAATGAGAAAAA ______ 2248 Right AACATGTCACCAACTGGGTATAACT 2269 Left AAAATCTCCAAAATATATGCCAAAG
2270 Right AACATGTCACCAACTGGGTATAACT
2249 Left AATCTCCAAAATATATGCCAAAGAA ______ 2250 Right AACATGTCACCAACTGGGTATAACT 2271 Left ATGCCAAAGAAGTAGAATGAGAAAA
2272 Right ACTGGGTATAACTGCACATTCAGAG
2251 Left AAATCTCCAAAATATATGCCAAAGA ______ 2252 Right AACATGTCACCAACTGGGTATAACT 2273 Left ATCTCCAAAATATATGCCAAAGAAG
2274 Right AACTGCACATTCAGAGATTCTTTCT
2253 Left TCTCCAAAATATATGCCAAAGAAGT ______ 2254 Right AACTGCACATTCAGAGATTCTTTCT 2275 Left AATCTCCAAAATATATGCCAAAGAAG
2276 Right AACATGTCACCAACTGGGTATAACT
2255 Left TCTCCAAAATATATGCCAAAGAAGT ______ 2256 Right GGGTATAACTGCACATTCAGAGATT 2277 Left ATCTCCAAAATATATGCCAAAGAAG
2278 Right GGGTATAACTGCACATTCAGAGATT
2257 Left TGCCAAAGAAGTAGAATGAGAAAAA ______ 2258 Right GGGTATAACTGCACATTCAGAGATT 2279 Left AAAAATCTCCAAAATATATGCCAAAG
2280 Right AACTGCACATTCAGAGATTCTTTCT
2259 Left AAATCTCCAAAATATATGCCAAAGA
2260 Right AACTGCACATTCAGAGATTCTTTCT
EGFR Exon20-21 2kb S eq. 2300 Right AAAGATGAGATAACTTGGTGGAGTG
ID Primer Sequence 2281 Left TTTCTACCAACTTCTGTCAAGCTCT 2301 Left AGAAAGAATCTCTGAATGTGCAGTT
2282 Right TGCTATGTATTCTGTGGGTTAGACA 2302 Right CAAAGATGAGATAACTTGGTGGAGT
2283 Left TTTCTACCAACTTCTGTCAAGCTCT 2303 Left AGAAAGAATCTCTGAATGTGCAGTT
2284 Right TGAGGTAATAAGTCAGCCATTTTTC 2304 Right TTTTCCAAGAACTTATTCCACAAAG
2285 Left TTTCTACCAACTTCTGTCAAGCTCT 2305 Left AATCTCTGAATGTGCAGTTATACCC
2286 Right GGTAATAAGTCAGCCATTTTTCCTT 2306 Right AAAGATGAGATAACTTGGTGGAGTG
2287 Left AGAAAGAATCTCTGAATGTGCAGTT 2307 Left AATCTCTGAATGTGCAGTTATACCC
2288 Right CAAAGTAACAATCAACAGACACTGG 2308 Right CAAAGATGAGATAACTTGGTGGAGT
2289 Left TTTCTACCAACTTCTGTCAAGCTCT
2290 Right ATAAAGGCCCATGTTCTCTTTACTT
2309 Left TTTCTACCAACTTCTGTCAAGCTCT
2291 Left TTTCTACCAACTTCTGTCAAGCTCT 2310 Right AATAAGTCAGCCATTTTTCCTTTTC
2292 Right CCTTCTTGGCTGTAAGATCAACTAA ____ 2311 Left TTTCTACCAACTTCTGTCAAGCTCT
2293 Left AATCTCTGAATGTGCAGTTATACCC 2312 Right CTGCCCAGAGAAAATTAAACTGTAG
2294 Right CAAAGTAACAATCAACAGACACTGG _____ 2313 Left AATCTCTGAATGTGCAGTTATACCC
2314 Right TTTTCCAAGAACTTATTCCACAAAG
S eq. 2315 Left GTTTCTACCAACTTCTGTCAAGCTC

Primer Sequence ID 2316 Right GGTAATAAGTCAGCCATTTTTCCTT
2295 Left TTTCTACCAACTTCTGTCAAGCTCT _____ 2296 Right ATTACTCTCTGGCTTTTGTCCTTCT 2317 Left TGTGTTTCTACCAACTTCTGTCAAG
2318 Right TGCTATGTATTCTGTGGGTTAGACA
2297 Left GTTTCTACCAACTTCTGTCAAGCTC _____ 2298 Right TGAGGTAATAAGTCAGCCATTTTTC 2319 Left GTTTCTACCAACTTCTGTCAAGCTC
2320 Right ATAAAGGCCCATGTTCTCTTTACTT
2299 Left AGAAAGAATCTCTGAATGTGCAGTT
EGFR Ex n22 2kb S eq. 2331 Left GTCTGTAGGTTACACACAAATGCTG

Primer Sequence ID 2332 Right TTCTCCACTACAAATCACCACAGTA
2321 Left CACATAGCATTTGCACTGTATTAGG _ ________________________________________________ 2322 Right TTGAATCCAAAATAAAGGAATGTGT 2333 Left AGGTAATCAGGAGATGCTGTAGATG
2334 Right CACACTGAGCACTCAATAAAGAGAA
2323 Left ATTTTGATATTTAAGGGAGGTCCTG
2324 Right TTCTCCACTACAAATCACCACAGTA 2335 Left GTCTGTAGGTTACACACAAATGCTG
2336 Right ATTCTTCAAAGGTAGCTGATTGATG
_ 2325 Left CACATAGCATTTGCACTGTATTAGG
2326 Right TTCTCCACTACAAATCACCACAGTA 2337 Left ATTTTGATATTTAAGGGAGGTCCTG
2338 Right GCTTACCTTGTTATCAAGTCCTGAA
2327 Left GTCTGTAGGTTACACACAAATGCTG
2328 Right CACACTGAGCACTCAATAAAGAGAA 2339 Left CACATAGCATTTGCACTGTATTAGG
2340 Right GCTTACCTTGTTATCAAGTCCTGAA
2329 Left CACATAGCATTTGCACTGTATTAGG
2330 Right ATTCTTCAAAGGTAGCTGATTGATG Seq.
ID Primer Sequence 2341 Left AGGTAATCAGGAGATGCTGTAGATG 2351 Left CTCTGAGAAAGAGTCTGCTAAGGAA
2342 Right ATTCTTCAAAGGTAGCTGATTGATG 2352 Right TTGAATCCAAAATAAAGGAATGTGT
2343 Left GTCTGTAGGTTACACACAAATGCTG 2353 Left TCGGTACTGAACATATACGGACTTT
2344 Right GCTTACCTTGTTATCAAGTCCTGAA 2354 Right TTGAATCCAAAATAAAGGAATGTGT
2345 Left AGGTAATCAGGAGATGCTGTAGATG 2355 Left TCGGTACTGAACATATACGGACTTT
2346 Right GCTTACCTTGTTATCAAGTCCTGAA 2356 Right CACACTGAGCACTCAATAAAGAGAA
2347 Left GTATTTTGAAACTCAAGATCGCATT 2357 Left CACATAGCATTTGCACTGTATTAGG
2348 Right TTATACACATAGCGGAGTGATCAAA 2358 Right CAAAGGTAGCTGATTGATGAGAGTT
2349 Left TCTGAGAAAGAGTCTGCTAAGGAAG 2359 Left TCTGAGAAAGAGTCTGCTAAGGAAG
2350 Right TTGAATCCAAAATAAAGGAATGTGT 2360 Right ATCCAAAATAAAGGAATGTGTGTGT
EGFR Exon18-21 5kb S eq. .
Primer S equence ID 2379 Left TGCCAAAGAAGTAGAATGAGAAAAA
2361 Left ATGCCAAAGAAGTAGAATGAGAAAA 2380 Right TGCTATGTATTCTGTGGGTTAGACA
2362 Right TGCTATGTATTCTGTGGGTTAGACA
2381 Left ATGCCAAAGAAGTAGAATGAGAAAA
2363 Left ATGCCAAAGAAGTAGAATGAGAAAA 2382 Right AGACATTTTTATAAAGGCCCATGTT
2364 Right CCTTCTTGGCTGTAAGATCAACTAA ____ 2383 Left ATGCCAAAGAAGTAGAATGAGAAAA
2384 Right ATTGTAAGTGAAAGGCTTCACAGAT
S eq. . 2385 Left TCTCCAAAATATATGCCAAAGAAGT
Primer Sequence ID 2386 Right GGTAATAAGTCAGCCATTTTTCCTT
2365 Left ATGCCAAAGAAGTAGAATGAGAAAA
2366 Right GTGCACTTAACTTTTAAGCCTTGAC 2387 Left AATCTCCAAAATATATGCCAAAGAA
2388 Right ATCTATCTTCTACCCCATTTCCAAC
2367 Left ATGCCAAAGAAGTAGAATGAGAAAA
2368 Right AGATTGTAAGTGAAAGGCTTCACAG 2389 Left AAATCTCCAAAATATATGCCAAAGA
2390 Right ATCTATCTTCTACCCCATTTCCAAC
2391 Left AAATCTCCAAAATATATGCCAAAGA
2369 Left TCTCCAAAATATATGCCAAAGAAGT 2392 Right TGCTATGTATTCTGTGGGTTAGACA
2370 Right ATCTATCTTCTACCCCATTTCCAAC _____ ¨2393 Left TCTCCAAAATATATGCCAAAGAAGT
2371 Left ATGCCAAAGAAGTAGAATGAGAAAA 2394 Right CCTTCTTGGCTGTAAGATCAACTAA
2372 Right CTGCCCAGAGAAAATTAAACTGTAG
__________________________________________ 2395 Left TCTCCAAAATATATGCCAAAGAAGT
2373 Left TCTCCAAAATATATGCCAAAGAAGT 2396 Right GTGCACTTAACTTTTAAGCCTTGAC
2374 Right TGCTATGTATTCTGTGGGTTAGACA
__________________________________________ 2397 Left AATCTCCAAAATATATGCCAAAGAA
2375 Left TGCCAAAGAAGTAGAATGAGAAAAA 2398 Right TGAGGTAATAAGTCAGCCATTTTTC
2376 Right ATCTATCTTCTACCCCATTTCCAAC ____ 4 399 Left AAATCTCCAAAATATATGCCAAAGA
2377 Left TCTCCAAAATATATGCCAAAGAAGT 400 Right TGAGGTAATAAGTCAGCCATTTTTC
2378 Right TGAGGTAATAAGTCAGCCATTTTTC
EGFR Exon22 5kb S eq. Primer Sequence ID

2401 Left GTTGGAAATGGGGTAGAAGATAGAT 2421 Left CACATAGCATTTGCACTGTATTAGG
2402 Right TTGAATCCAAAATAAAGGAATGTGT 2422 Right GGGTCAAATAAACCTCCACTTATCT
2403 Left GTTGGAAATGGGGTAGAAGATAGAT 2423 Left GTTGGAAATGGGGTAGAAGATAGAT
2404 Right CACACTGAGCACTCAATAAAGAGAA 2424 Right ATTCTTCAAAGGTAGCTGATTGATG
2405 Left ATTTTGATATTTAAGGGAGGTCCTG 2425 Left ATTTTGATATTTAAGGGAGGTCCTG
2406 Right CTCTCCCATCAACATTTAGAAGAAA 2426 Right TATAAGCCAATAAATCCCATTTTGA
2407 Left CACATAGCATTTGCACTGTATTAGG 2427 Left CACATAGCATTTGCACTGTATTAGG
2408 Right CTCTCCCATCAACATTTAGAAGAAA 2428 Right TATAAGCCAATAAATCCCATTTTGA
2409 Left ATTTTGATATTTAAGGGAGGTCCTG 2429 Left GTCTGTAGGTTACACACAAATGCTG
2410 Right TACAACAAACACAAGAATGGCTTTA 2430 Right TACAACAAACACAAGAATGGCTTTA
2411 Left CACATAGCATTTGCACTGTATTAGG 2431 Left TACAGATTATGATGACTGCCTCAAA
2412 Right TACAACAAACACAAGAATGGCTTTA 2432 Right TACAACAAACACAAGAATGGCTTTA
2413 Left GTTGGAAATGGGGTAGAAGATAGAT 2433 Left AGGAAAATAACACACACTCTCCTTG
2414 Right TTCTCCACTACAAATCACCACAGTA 2434 Right TTACTGGGAGATGATTAAGAACAGC
2415 Left ATATCTGAATAAAAGGTCACCAC CA 2435 Left GTCTGTAGGTTACACACAAATGCTG
2416 Right CTCTCCCATCAACATTTAGAAGAAA 2436 Right GGGTCAAATAAACCTCCACTTATCT
2417 Left CCATATCTGAATAAAAGGTCACCAC 2437 Left ATATCTGAATAAAAGGTCACCACCA
2418 Right CTCTCCCATCAACATTTAGAAGAAA 2438 Right TATAAGCCAATAAATCCCATTTTGA
S eq. .
Primer S equence ID 2439 Left CCATATCTGAATAAAAGGTCACCAC

2419 Left TATCTGAATAAAAGGTCACCACCAT 2440 Right TATAAGCCAATAAATCCCATTTTGA
2420 Right CTCTCCCATCAACATTTAGAAGAAA
Table 9. KIT Capture Primer List for NGS Panel KIT Exon8 150-175 bases S eq. . 2452 Right ATAAGCAGTGCCAAAAATAATCATC
ID Pnmer Sequence 2441 Left ATATGGCCATTTCTGTTTTCCTGTA 2453 Left TGACATATGGCCATTTCTGTTTT
2442 Right ATAAGCAGTGCCAAAAATAATCATC 2454 Right ATAAGCAGTGCCAAAAATAATCATC
2443 Left CTGACATATGGCCATTTCTGTTT 2455 Left ATATGGCCATTTCTGTTTTCCTGTA
2444 Right ATAAGCAGTGCCAAAAATAATCATC 2456 Right TTATAAGCAGTGCCAAAAATAATCA
2445 Left GACATATGGCCATTTCTGTTTTC 72457 Left TATGGCCATTTCTGTTTTCCTGTA
2446 Right ATAAGCAGTGCCAAAAATAATCATC 2458 Right ATAAGCAGTGCCAAAAATAATCATC
2447 Left CTGACATATGGCCATTTCTGTTTT 2459 Left GACATATGGCCATTTCTGTTTTC
2448 Right ATAAGCAGTGCCAAAAATAATCATC 2460 Right TTATAAGCAGTGCCAAAAATAATCA
2449 Left ATATGGCCATTTCTGTTTTCCTGTA Seq. .
S
2450 Right GCATTATAAGCAGTGCCAAAAATAA ID Primer equence 2461 Left CTGACATATGGCCATTTCTGTTTTC
2451 Left TATGGCCATTTCTGTTTTCCTGTAG 2462 Right ATAAGCAGTGCCAAAAATAATCATC

2463 Left ATATGGCCATTTCTGTTTTCCTGTA 2473 Left TATGGCCATTTCTGTTTTCCTGTAG
2464 Right TATAAGCAGTGCCAAAAATAATCATC 2474 Right TTATAAGCAGTGCCAAAAATAATCA
2465 Left GGCCATTTCTGTTTTCCTGTAG 2475 Left TGACATATGGCCATTTCTGTTTTC
2466 Right ATAAGCAGTGCCAAAAATAATCATC 2476 Right ATAAGCAGTGCCAAAAATAATCATC
2467 Left TATGGCCATTTCTGTTTTCCTGTAG 2477 Left ATATGGCCATTTCTGTTTTCCTGTA
2468 Right GCATTATAAGCAGTGCCAAAAATAA 2478 Right TAAGCAGTGCCAAAAATAATCATC
2469 Left ATATGGCCATTTCTGTTTTCCTGT 2479 Left GACATATGGCCATTTCTGTTTTC
2470 Right ATAAGCAGTGCCAAAAATAATCATC 2480 Right TATAAGCAGTGCCAAAAATAATCATC
2471 Left ACATATGGCCATTTCTGTTTTCCT
2472 Right ATAAGCAGTGCCAAAAATAATCATC
KIT Exon8 176-200 bases S eq. .
Pnmer Sequence ID 2501 Left GACATATGGCCATTTCTGTTTTC
2481 Left AGGTTTTCCAGCACTCTGACATA 2502 Right GCATTATAAGCAGTGCCAAAAATAA
2482 Right ATAAGCAGTGCCAAAAATAATCATC
2503 Left AGGTTTTCCAGCACTCTGACATA
2483 Left ACTCTGACATATGGCCATTTCTGTT 2504 Right TTATAAGCAGTGCCAAAAATAATCA
2484 Right ATAAGCAGTGCCAAAAATAATCATC
2505 Left ACTCTGACATATGGCCATTTCTGTT
2485 Left CTGACATATGGCCATTTCTGTTT 2506 Right TTATAAGCAGTGCCAAAAATAATCA
2486 Right ATAAGCAGTGCCAAAAATAATCATC ____ 2507 Left GAGGTTTTCCAGCACTCTGACATA
2487 Left CTGACATATGGCCATTTCTGTTTT 2508 Right ATAAGCAGTGCCAAAAATAATCATC
2488 Right ATAAGCAGTGCCAAAAATAATCATC
S eq. . 2509 Left CTGACATATGGCCATTTCTGTTT
Pnmer Sequence ID 2510 Right TTATAAGCAGTGCCAAAAATAATCA
2489 Left ATATGGCCATTTCTGTTTTCCTGTA _____ 2490 Right GCATTATAAGCAGTGCCAAAAATAA 2511 Left TCTGACATATGGCCATTTCTGTTT
2512 Right ATAAGCAGTGCCAAAAATAATCATC
2491 Left CTCTGACATATGGCCATTTCTGTT ______ 2492 Right ATAAGCAGTGCCAAAAATAATCATC 2513 Left GACATATGGCCATTTCTGTTTTC
2514 Right TTATAAGCAGTGCCAAAAATAATCA
2493 Left CTCTGACATATGGCCATTTCTGTTT
2494 Right ATAAGCAGTGCCAAAAATAATCATC 2515 Left CTGACATATGGCCATTTCTGTTTTC
2516 Right ATAAGCAGTGCCAAAAATAATCATC
2495 Left AGGTTTTCCAGCACTCTGACATA _______ 2496 Right GCATTATAAGCAGTGCCAAAAATAA 2517 Left TCTGACATATGGCCATTTCTGTTTT
2518 Right ATAAGCAGTGCCAAAAATAATCATC
2497 Left ACTCTGACATATGGCCATTTCTGTT
2498 Right GCATTATAAGCAGTGCCAAAAATAA 2519 Left CTGACATATGGCCATTTCTGTTTT
2520 Right TTATAAGCAGTGCCAAAAATAATCA
2499 Left TCTGACATATGGCCATTTCTGTT
2500 Right ATAAGCAGTGCCAAAAATAATCATC
KIT Exon8 201-300 bases Seq. 2522 Right ATAAGCAGTGCCAAAAATAATCATC
ID Primer Sequence 2521 Left TTAGAGAGGGAGTGAAGTGAATGTT 2523 Left GATTAGAGAGGGAGTGAAGTGAATG

2524 Right ATAAGCAGTGCCAAAAATAATCATC 2542 Right TTATAAGCAGTGCCAAAAATAATCA
2525 Left GGATTAGAGAGGGAGTGAAGTGAAT 2543 Left GGATTAGAGAGGGAGTGAAGTGAAT
2526 Right ATAAGCAGTGCCAAAAATAATCATC 2544 Right TTATAAGCAGTGCCAAAAATAATCA
2527 Left GTAGGGATTAGAGAGGGAGTGAAGT 2545 Left TCAGGAAGGTTGTAGGGATTAGAG
2528 Right ATAAGCAGTGCCAAAAATAATCATC 2546 Right ATAAGCAGTGCCAAAAATAATCATC
2529 Left TTAGAGAGGGAGTGAAGTGAATGTT 2547 Left CAGGAAGGTTGTAGGGATTAGAGA
2530 Right GCATTATAAGCAGTGCCAAAAATAA 2548 Right ATAAGCAGTGCCAAAAATAATCATC
2531 Left GATTAGAGAGGGAGTGAAGTGAATG 2549 Left CTCAGGAAGGTTGTAGGGATTAGA
2532 Right GCATTATAAGCAGTGCCAAAAATAA 2550 Right ATAAGCAGTGCCAAAAATAATCATC
2533 Left CAGGAAGGTTGTAGGGATTAGAGAG 2551 Left TAGAGAGGGAGTGAAGTGAATGTTG
2534 Right ATAAGCAGTGCCAAAAATAATCATC 2552 Right ATAAGCAGTGCCAAAAATAATCATC
2535 Left CTCAGGAAGGTTGTAGGGATTAGAG 2553 Left GTAGGGATTAGAGAGGGAGTGAAGT
2536 Right ATAAGCAGTGCCAAAAATAATCATC 2554 Right TTATAAGCAGTGCCAAAAATAATCA
2537 Left GGATTAGAGAGGGAGTGAAGTGAAT 2555 Left AGGTTGTAGGGATTAGAGAGGGAGT
2538 Right GCATTATAAGCAGTGCCAAAAATAA 2556 Right ATAAGCAGTGCCAAAAATAATCATC
2539 Left GTAGGGATTAGAGAGGGAGTGAAGT 2557 Left ATTAGAGAGGGAGTGAAGTGAATGTT
2540 Right GCATTATAAGCAGTGCCAAAAATAA 2558 Right ATAAGCAGTGCCAAAAATAATCATC
Seq. . 2559 Left GGATTAGAGAGGGAGTGAAGTGAA
Pnmer Sequence ID 2560 Right ATAAGCAGTGCCAAAAATAATCATC
2541 Left GATTAGAGAGGGAGTGAAGTGAATG
KIT Exon8 301-400 bases Seq. . 2575 Left GATTAGAGAGGGAGTGAAGTGAATG

Pnmer Sequence ID 2576 Right AATTATCCCTTCTAAAAAGCCACAT
2561 Left TTAGAGAGGGAGTGAAGTGAATGTT
2562 Right TCATTCAGTAATGATTTTTCAGCAA 2577 Left GGATTAGAGAGGGAGTGAAGTGAAT
2578 Right AATTATCCCTTCTAAAAAGCCACAT
2563 Left TTAGAGAGGGAGTGAAGTGAATGTT
2564 Right TCAGTAATGATTTTTCAGCAAACAA 2579 Left TTAGAGAGGGAGTGAAGTGAATGTT
2580 Right GTCATTCAGTAATGATTTTTCAGCA
2565 Left GATTAGAGAGGGAGTGAAGTGAATG
2566 Right TCAGTAATGATTTTTCAGCAAACAA Seq. .
ID Primer Sequence 2567 Left GATTAGAGAGGGAGTGAAGTGAATG 2581 Left TTAGAGAGGGAGTGAAGTGAATGTT
2568 Right TTCAGTAATGATTTTTCAGCAAACA 2582 Right CAGCAAACAAAATTAATGTCTACCA
2569 Left GGATTAGAGAGGGAGTGAAGTGAAT 2583 Left GATTAGAGAGGGAGTGAAGTGAATG
2570 Right TCAGTAATGATTTTTCAGCAAACAA 2584 Right CAGCAAACAAAATTAATGTCTACCA
2571 Left GGATTAGAGAGGGAGTGAAGTGAAT 2585 Left GGATTAGAGAGGGAGTGAAGTGAAT
2572 Right TTCAGTAATGATTTTTCAGCAAACA 2586 Right CAGCAAACAAAATTAATGTCTACCA
2573 Left TTAGAGAGGGAGTGAAGTGAATGTT 2587 Left GTAGGGATTAGAGAGGGAGTGAAGT
2574 Right AATTATCCCTTCTAAAAAGCCACAT 2588 Right CAGCAAACAAAATTAATGTCTACCA

2589 Left TAGAGAGGGAGTGAAGTGAATGTTG 2595 Left TAGAGAGGGAGTGAAGTGAATGTTG
2590 Right TCATTCAGTAATGATTTTTCAGCAA 2596 Right TTCAGTAATGATTTTTCAGCAAACA
2591 Left ATTAGAGAGGGAGTGAAGTGAATGTT 2597 Left ATTAGAGAGGGAGTGAAGTGAATGTT
2592 Right TCATTCAGTAATGATTTTTCAGCAA 2598 Right TCAGTAATGATTTTTCAGCAAACAA
2593 Left TAGAGAGGGAGTGAAGTGAATGTTG 2599 Left CAGGAAGGTTGTAGGGATTAGAGAG
2594 Right TCAGTAATGATTTTTCAGCAAACAA 2600 Right AATTATCCCTTCTAAAAAGCCACAT
KIT Exon8 401-500 bases S eq. 2620 Right AAATTGCATGATAAATCCAGAAAGA
ID Pnmer Sequence 2601 Left GATTAGAGAGGGAGTGAAGTGAATG 2621 Left GGATTAGAGAGGGAGTGAAGTGAAT
2602 Right TCATTCAGTAATGATTTTTCAGCAA 2622 Right GAAATTGCATGATAAATCCAGAAAG
2603 Left GGATTAGAGAGGGAGTGAAGTGAAT ¨ 623 Left GTAGGGATTAGAGAGGGAGTGAAGT
2604 Right TCATTCAGTAATGATTTTTCAGCAA 2624 Right AAATTGCATGATAAATCCAGAAAGA
2605 Left GTAGGGATTAGAGAGGGAGTGAAGT 2625 Left GTAGGGATTAGAGAGGGAGTGAAGT
2606 Right TCATTCAGTAATGATTTTTCAGCAA 2626 Right GAAATTGCATGATAAATCCAGAAAG
2607 Left GTAGGGATTAGAGAGGGAGTGAAGT 2627 Left TTAGAAGCAGTCTTCAGATCCCTAC
2608 Right TCAGTAATGATTTTTCAGCAAACAA 2628 Right ATAAGCAGTGCCAAAAATAATCATC
2609 Left GTAGGGATTAGAGAGGGAGTGAAGT 2629 Left GATTAGAGAGGGAGTGAAGTGAATG
2610 Right TTCAGTAATGATTTTTCAGCAAACA 2630 Right GTCATTCAGTAATGATTTTTCAGCA
2611 Left TTAGAGAGGGAGTGAAGTGAATGTT 2631 Left GGATTAGAGAGGGAGTGAAGTGAAT
2612 Right AAATTGCATGATAAATCCAGAAAGA 2632 Right GTCATTCAGTAATGATTTTTCAGCA
S eq.
.633 Left AGATTTTTACCTGTGGAACACTTTG
Pnmer S equence ID 634 Right GCATTATAAGCAGTGCCAAAAATAA
2613 Left TTAGAGAGGGAGTGAAGTGAATGTT
2614 Right GAAATTGCATGATAAATCCAGAAAG 2635 Left GTAGGGATTAGAGAGGGAGTGAAGT
_______________________________________________________________________________ ____ 636 Right GTCATTCAGTAATGATTTTTCAGCA
2615 Left GATTAGAGAGGGAGTGAAGTGAATG ______ 2616 Right AAATTGCATGATAAATCCAGAAAGA 2637 Left CAGGAAGGTTGTAGGGATTAGAGAG
_______________________________________________________________________________ ____ 638 Right TCATTCAGTAATGATTTTTCAGCAA
2617 Left GATTAGAGAGGGAGTGAAGTGAATG ______ 2618 Right GAAATTGCATGATAAATCCAGAAAG 2639 Left CTCAGGAAGGTTGTAGGGATTAGAG
_______________________________________________________________________________ ____ 640 Right TCATTCAGTAATGATTTTTCAGCAA
2619 Left GGATTAGAGAGGGAGTGAAGTGAAT
KIT Exon8 501-600 bases S eq.
2646 Right CGATCATTACTTTTTGGTAACTTGG
ID Pnmer Sequence 2641 Left TTAGAGAGGGAGTGAAGTGAATGTT 2647 Left TTAGAGAGGGAGTGAAGTGAATGTT
2642 Right CGATCATTACTTTTTGGTAACTTGG 2648 Right CATTACTTTTTGGTAACTTGGCAAT
2643 Left GATTAGAGAGGGAGTGAAGTGAATG ¨ 649 Left TTAGAGAGGGAGTGAAGTGAATGTT
2644 Right CGATCATTACTTTTTGGTAACTTGG 2650 Right GCGATCATTACTTTTTGGTAACTTG
2645 Left GGATTAGAGAGGGAGTGAAGTGAAT 2651 Left TTAGAGAGGGAGTGAAGTGAATGTT

2652 Right TGCGATCATTACTTTTTGGTAACTT 2666 Right ATAAGCAGTGCCAAAAATAATCATC
2653 Left GATTAGAGAGGGAGTGAAGTGAATG 2667 Left TTAGAAGCAGTCTTCAGATCCCTAC
2654 Right CATTACTTTTTGGTAACTTGGCAAT 2668 Right AATTATCCCTTCTAAAAAGCCACAT
2655 Left GATTAGAGAGGGAGTGAAGTGAATG 2669 Left TTAGAGAGGGAGTGAAGTGAATGTT
2656 Right GCGATCATTACTTTTTGGTAACTTG 2670 Right GATAACTACAGTCACATTTCCCACA
2657 Left GGATTAGAGAGGGAGTGAAGTGAAT 2671 Left AACAACTCCTAATTTCATCCATTCC
2658 Right CATTACTTTTTGGTAACTTGGCAAT 2672 Right ATAAGCAGTGCCAAAAATAATCATC
2659 Left ACTCCTAATTTCATCCATTCCAGTT 2673 Left GATTAGAGAGGGAGTGAAGTGAATG
2660 Right ATAAGCAGTGCCAAAAATAATCATC 2674 Right GATAACTACAGTCACATTTCCCACA
S eq. . 2675 Left TTAGAGAGGGAGTGAAGTGAATGTT

Pnmer S equence ID 2676 Right TAACTACAGTCACATTTCCCACACA
2661 Left AACTCCTAATTTCATCCATTCCAGT
2662 Right ATAAGCAGTGCCAAAAATAATCATC 2677 Left GGATTAGAGAGGGAGTGAAGTGAAT
2678 Right GATAACTACAGTCACATTTCCCACA
2663 Left GTAGGGATTAGAGAGGGAGTGAAGT
2664 Right CATTACTTTTTGGTAACTTGGCAAT 2679 Left CACTTTGGAGTCCTAGAGTTTGATT
2680 Right AATTATCCCTTCTAAAAAGCCACAT
2665 Left GGCAGGAATCCTTTAAAGTAGATTT
KIT Exon8 601-800 bases S eq. .
Primer S equence ID 2699 Left GGGCTTATCTTTTCCTCTAACAACT

2681 Left AGATTTTTACCTGTGGAACACTTTG 2700 Right TCATTCAGTAATGATTTTTCAGCAA
2682 Right TCATTCAGTAATGATTTTTCAGCAA
2701 Left GGGCTTATCTTTTCCTCTAACAACT
2683 Left AGATTTTTACCTGTGGAACACTTTG 2702 Right TCAGTAATGATTTTTCAGCAAACAA
2684 Right TTCAGTAATGATTTTTCAGCAAACA ___ S eq. . 2703 Left GGGCTTATCTTTTCCTCTAACAACT

Pnmer S equence ID 2704 Right TTCAGTAATGATTTTTCAGCAAACA
2685 Left AGATTTTTACCTGTGGAACACTTTG _____ 2686 Right TCAGTAATGATTTTTCAGCAAACAA 2705 Left AGATTTTTACCTGTGGAACACTTTG
2706 Right AATTATCCCTTCTAAAAAGCCACAT
2687 Left GGAGAAAATTCATGTAAGAGCAAAA ______ 2688 Right ATAAGCAGTGCCAAAAATAATCATC 2707 Left AGGCTGGTTTTCTTTTCTAGTTTTC
2708 Right ATAAGCAGTGCCAAAAATAATCATC
2689 Left AATTCATGTAAGAGCAAAAGAGTGG ______ 2690 Right ATAAGCAGTGCCAAAAATAATCATC 2709 Left GGCTGGTTTTCTTTTCTAGTTTTCT
2710 Right ATAAGCAGTGCCAAAAATAATCATC
2691 Left GGAGAAAATTCATGTAAGAGCAAAA ______ 2692 Right AATTATCCCTTCTAAAAAGCCACAT 2711 Left ACTCCTAATTTCATCCATTCCAGTT
2712 Right TCATTCAGTAATGATTTTTCAGCAA
2693 Left AGATTTTTACCTGTGGAACACTTTG _____ 2694 Right AAATTGCATGATAAATCCAGAAAGA 2713 Left AACTCCTAATTTCATCCATTCCAGT
2714 Right TCATTCAGTAATGATTTTTCAGCAA
2695 Left AGATTTTTACCTGTGGAACACTTTG _____ 2696 Right GAAATTGCATGATAAATCCAGAAAG 2715 Left AGGAGAAAATTCATGTAAGAGCAAA
2716 Right ATAAGCAGTGCCAAAAATAATCATC
2697 Left AATTCATGTAAGAGCAAAAGAGTGG ______ 2698 Right AATTATCCCTTCTAAAAAGCCACAT 717 Left AAGGAGAAAATTCATGTAAGAGCAA

2718 Right ATAAGCAGTGCCAAAAATAATCATC 2719 Left AAAGGAGAAAATTCATGTAAGAGCA
2720 Right ATAAGCAGTGCCAAAAATAATCATC
KIT Exon8 801-1000 bases S eq. .
Primer S equence ID 2741 Left AGGCTGGTTTTCTTTTCTAGTTTTC

2721 Left GGAGAAAATTCATGTAAGAGCAAAA 2742 Right TCATTCAGTAATGATTTTTCAGCAA
2722 Right TCATTCAGTAATGATTTTTCAGCAA
2743 Left GGCTGGTTTTCTTTTCTAGTTTTCT
2723 Left GGAGAAAATTCATGTAAGAGCAAAA 2744 Right TCATTCAGTAATGATTTTTCAGCAA
2724 Right TCAGTAATGATTTTTCAGCAAACAA
2745 Left AATTCATGTAAGAGCAAAAGAGTGG
2725 Left GGAGAAAATTCATGTAAGAGCAAAA 2746 Right AAATTGCATGATAAATCCAGAAAGA
2726 Right TTCAGTAATGATTTTTCAGCAAACA
2747 Left AATTCATGTAAGAGCAAAAGAGTGG
2727 Left AATTCATGTAAGAGCAAAAGAGTGG 2748 Right GAAATTGCATGATAAATCCAGAAAG
2728 Right TCATTCAGTAATGATTTTTCAGCAA
2749 Left AGATTTTTACCTGTGGAACACTTTG
2729 Left AATTCATGTAAGAGCAAAAGAGTGG 2750 Right TGTTGTAATTGTGCGATCATTACTT
2730 Right TCAGTAATGATTTTTCAGCAAACAA
2751 Left AGGCTGGTTTTCTTTTCTAGTTTTC
2731 Left AATTCATGTAAGAGCAAAAGAGTGG 2752 Right TTCAGTAATGATTTTTCAGCAAACA
2732 Right TTCAGTAATGATTTTTCAGCAAACA
2753 Left AGATTTTTACCTGTGGAACACTTTG
2733 Left TTAGAGCATTTCTGCTGTTACAGTG 2754 Right CGATCATTACTTTTTGGTAACTTGG
2734 Right ATAAGCAGTGCCAAAAATAATCATC
2755 Left TTAGAGAGGGAGTGAAGTGAATGTT
2735 Left GGAGAAAATTCATGTAAGAGCAAAA 2756 Right GAACCCTACTTAGTATTCCCCAAAA
2736 Right AAATTGCATGATAAATCCAGAAAGA
_________________________________________ 2757 Left AGGAGAAAATTCATGTAAGAGCAAA

2737 Left GGAGAAAATTCATGTAAGAGCAAAA 2758 Right TCATTCAGTAATGATTTTTCAGCAA
2738 Right GAAATTGCATGATAAATCCAGAAAG ____ S eq. . 2759 Left AAGGAGAAAATTCATGTAAGAGCAA

Pnmer S equence ID 2760 Right TCATTCAGTAATGATTTTTCAGCAA
2739 Left ATACCAAATTAGAGCATTTCTGCTG
2740 Right ATAAGCAGTGCCAAAAATAATCATC
KIT Exon8 2kb S eq. .
Primer S equence ID 2771 Left AATTCATGTAAGAGCAAAAGAGTGG

2761 Left GGAGAAAATTCATGTAAGAGCAAAA 2772 Right GGACTGAAGTTTGAGTTCTAAGCAG
2762 Right GGACTGAAGTTTGAGTTCTAAGCAG
2773 Left AATTCATGTAAGAGCAAAAGAGTGG
2763 Left GGAGAAAATTCATGTAAGAGCAAAA 2774 Right AGGACTGAAGTTTGAGTTCTAAGCA
2764 Right AGGACTGAAGTTTGAGTTCTAAGCA
2775 Left GGAGAAAATTCATGTAAGAGCAAAA
2765 Left TTAGAGCATTTCTGCTGTTACAGTG 2776 Right GACCAGAAAATAGTCAAAGTGAGGA
2766 Right CCTGTTTCCTTTCTTAACACCTACA
_________________________________________ 2777 Left TTCTGATCTGTCAGTCTTTCCTTCT

2767 Left GGAGAAAATTCATGTAAGAGCAAAA 2778 Right TCATTCAGTAATGATTTTTCAGCAA
2768 Right CCTGTTTCCTTTCTTAACACCTACA ____ 2779 Left ATACCAAATTAGAGCATTTCTGCTG
2769 Left AATTCATGTAAGAGCAAAAGAGTGG 2780 Right CCTGTTTCCTTTCTTAACACCTACA
2770 Right ACTGAAGTTTGAGTTCTAAGCAGGA

S eq.
Primer S equence ID 2791 Left AATTCATGTAAGAGCAAAAGAGTGG

2781 Left TTAGAGCATTTCTGCTGTTACAGTG 2792 Right GACCAGAAAATAGTCAAAGTGAGGA
2782 Right GCTATTCCCTGTTTCCTTTCTTAAC
2793 Left TTCCTTCTCACTGCATATATTTTCC
_ 2783 Left ATAAGTGCATCTTCCTTTCACTTTG 2794 Right TCATTCAGTAATGATTTTTCAGCAA
2784 Right GAACCCTACTTAGTATTCCCCAAAA
_________________________________________ 2795 Left ATACCAAATTAGAGCATTTCTGCTG

_ 2785 Left GGAGAAAATTCATGTAAGAGCAAAA 2796 Right GCTATTCCCTGTTTCCTTTCTTAAC
2786 Right GCTATTCCCTGTTTCCTTTCTTAAC _ ________________________________________________ 2797 Left TGAAATTGGTTATCCAAGAAAGGTA
2787 Left TTCTGATCTGTCAGTCTTTCCTTCT 2798 Right TCATTCAGTAATGATTTTTCAGCAA
2788 Right TTCAGTAATGATTTTTCAGCAAACA
2799 Left AGATTTTTACCTGTGGAACACTTTG
2789 Left TTCTGATCTGTCAGTCTTTCCTTCT 2800 Right ACTGAAGTTTGAGTTCTAAGCAGGA
2790 Right TCAGTAATGATTTTTCAGCAAACAA
KIT Exon8-9 5kb S eq.
Primer S equence ID 2821 Left AGTTGCCCATGATAATTAAATGAAA

2801 Left AGTTGCCCATGATAATTAAATGAAA 2822 Right GCTTCCTTTATGGACGGTTTATATT
2802 Right AGGCAGTGTTAACTTTTGGATACAG
2823 Left CCCATGATAATTAAATGAAACTTGC
2803 Left CCCATGATAATTAAATGAAACTTGC 2824 Right GCTTCCTTTATGGACGGTTTATATT
2804 Right AGGCAGTGTTAACTTTTGGATACAG
2825 Left TGCCCATGATAATTAAATGAAACTT
2805 Left GCCCATGATAATTAAATGAAACTTG 2826 Right GCTTCCTTTATGGACGGTTTATATT
2806 Right AGGCAGTGTTAACTTTTGGATACAG
2827 Left TTGCCCATGATAATTAAATGAAACT
2807 Left TGCCCATGATAATTAAATGAAACTT 2828 Right CCCCTTAAATTGGATTAAAAAGAAA
2808 Right AGGCAGTGTTAACTTTTGGATACAG
_________________________________________ 2829 Left AGTTGCCCATGATAATTAAATGAAA

_ 2809 Left TTGCCCATGATAATTAAATGAAACT 2830 Right CCCCTTAAATTGGATTAAAAAGAAA
2810 Right AGGCAGTGTTAACTTTTGGATACAG
S eq.2831 Left GCCCATGATAATTAAATGAAACTTG
Primer S equence ID 2832 Right CCCCTTAAATTGGATTAAAAAGAAA
2811 Left AGTTTAGGCTTGCTTAGAAAAGGAG ______ 2812 Right AGGCAGTGTTAACTTTTGGATACAG 2833 Left CCCATGATAATTAAATGAAACTTGC
2834 Right CCCCTTAAATTGGATTAAAAAGAAA
2813 Left AGAGTTTAGGCTTGCTTAGAAAAGG ______ 2814 Right AGGCAGTGTTAACTTTTGGATACAG 2835 Left TGCCCATGATAATTAAATGAAACTT
2836 Right CCCCTTAAATTGGATTAAAAAGAAA
2815 Left GATTTCTTGTGTCGTGTCCTACTTT _____ 2816 Right AGGCAGTGTTAACTTTTGGATACAG 2837 Left AGTTTAGGCTTGCTTAGAAAAGGAG
2838 Right CCCCTTAAATTGGATTAAAAAGAAA
2817 Left GCCCATGATAATTAAATGAAACTTG _____ 2818 Right GCTTCCTTTATGGACGGTTTATATT 2839 Left AGAGTTTAGGCTTGCTTAGAAAAGG
2840 Right CCCCTTAAATTGGATTAAAAAGAAA
2819 Left TTGCCCATGATAATTAAATGAAACT
2820 Right GCTTCCTTTATGGACGGTTTATATT
KIT Exon9 200-250 bases Seq..2841 Left GGCTTTTGTTTTCTTCCCTTTAG
Prmer Sequence ID 2842 Right CATCCCCTTAAATTGGATTAAAAAG

ID
2843 Left GGCTTTTGTTTTCTTCCCTTTAG 2861 Left AGGGCTTTTGTTTTCTTCCCTTTAG
2844 Right ATCCCCTTAAATTGGATTAAAAAG 2862 Right CCCCTTAAATTGGATTAAAAAG
2845 Left GGGCTTTTGTTTTCTTCCCTTTAG 2863 Left GGGCTTTTGTTTTCTTCCCTTTAG
2846 Right ATCCCCTTAAATTGGATTAAAAAG 2864 Right CCCCTTAAATTGGATTAAAAAG
2847 Left GGCTTTTGTTTTCTTCCCTTTAG 2865 Left GCTTTTGTTTTCTTCCCTTTAG
2848 Right TCCCCTTAAATTGGATTAAAAAG 2866 Right TCCCCTTAAATTGGATTAAAAAG
2849 Left GCTTTTGTTTTCTTCCCTTTAG 2867 Left CTTTTGTTTTCTTCCCTTTAG
2850 Right CATCCCCTTAAATTGGATTAAAAAG 2868 Right CATCCCCTTAAATTGGATTAAAAAG
2851 Left AGGGCTTTTGTTTTCTTCCCTTTAG 2869 Left CAGGGCTTTTGTTTTCTTCC
2852 Right TCCCCTTAAATTGGATTAAAAAG CC
CCTTAAATTGGATTAAAAAGAAAT
Right 2853 Left GGGCTTTTGTTTTCTTCCCTTTAG
2854 Right TCCCCTTAAATTGGATTAAAAAG 2871 Left CTTTTGTTTTCTTCCCTTTAG
2872 Right ACATCCCCTTAAATTGGATTAAAAAG
2855 Left GCTTTTGTTTTCTTCCCTTTAG
2856 Right ACATCCCCTTAAATTGGATTAAAAAG 2873 Left CAGGGCTTTTGTTTTCTTCCCTTTAG
2874 Right CCCCTTAAATTGGATTAAAAAG
2857 Left GGCTTTTGTTTTCTTCCCTTTAG
2858 Right CCCCTTAAATTGGATTAAAAAG 2875 Left CAGGGCTTTTGTTTTCTTCC
2876 Right CCCCTTAAATTGGATTAAAAAG
2859 Left GCTTTTGTTTTCTTCCCTTTAG
2860 Right ATCCCCTTAAATTGGATTAAAAAG 2877 Left GGCTTTTGTTTTCTTCCCTTTAG
2878 Right CATCCCCTTAAATTGGATTA
2879 Left GCTTTTGTTTTCTTCCCTTTAG
2880 Right CCCCTTAAATTGGATTAAAAAG
Seq. Primer Sequence KIT Exon9 251-300 bases S eq. . 2892 Right ATCCCCTTAAATTGGATTAAAAAGAA
ID Primer Sequence 2881 Left CCACATCCCAAGTGTTTTATGTATT 2893 Left CCACATCCCAAGTGTTTTATGTAT
2882 Right CCCCTTAAATTGGATTAAAAAGAAA 2894 Right CCCCTTAAATTGGATTAAAAAGAAA
2883 Left CCACATCCCAAGTGTTTTATGTATT 2895 Left CCACATCCCAAGTGTTTTATGTATT
2884 Right CATCCCCTTAAATTGGATTAAAAAG 2896 Right TCCCCTTAAATTGGATTAAAAAGA
2885 Left CCACATCCCAAGTGTTTTATGTATT 2897 Left CCACATCCCAAGTGTTTTATGTAT
2886 Right TCCCCTTAAATTGGATTAAAAAGAA 2898 Right CATCCCCTTAAATTGGATTAAAAAG
2887 Left CCACATCCCAAGTGTTTTATGTATT 2899 Left CCACATCCCAAGTGTTTTATGTAT
2888 Right ATCCCCTTAAATTGGATTAAAAAGA 2900 Right TCCCCTTAAATTGGATTAAAAAGAA
2889 Left CCACATCCCAAGTGTTTTATGTATT Seq. .
2890 Right CCCCTTAAATTGGATTAAAAAGAAAT ID Pnmer Sequence 2901 Left GCCACATCCCAAGTGTTTTATGTAT
2891 Left CCACATCCCAAGTGTTTTATGTATT 2902 Right CCCCTTAAATTGGATTAAAAAGAAA

2903 Left CCACATCCCAAGTGTTTTATGTAT 2913 Left CTAGAGTAAGCCAGGGCTTTTGTTT
2904 Right ATCCCCTTAAATTGGATTAAAAAGA 2914 Right AACATCCCCTTAAATTGGATTAAAA
2905 Left CCACATCCCAAGTGTTTTATGTATT 2915 Left CTAGAGTAAGCCAGGGCTTTTGTT
2906 Right CCCCTTAAATTGGATTAAAAAGAA 2916 Right AACATCCCCTTAAATTGGATTAAAA
2907 Left GGCTTTTGTTTTCTTCCCTTTAG 2917 Left CCACATCCCAAGTGTTTTATGTATT
2908 Right TATGGTAGACAGAGCCTAAACATCC 2918 Right TCCCCTTAAATTGGATTAAAAAGAAA
2909 Left CTAGAGTAAGCCAGGGCTTTTGTTT 2919 Left GGCTTTTGTTTTCTTCCCTTTAG
2910 Right TATGGTAGACAGAGCCTAAACATCC 2920 Right TCATGACTGATATGGTAGACAGAGC
2911 Left CTAGAGTAAGCCAGGGCTTTTGTT
2912 Right TATGGTAGACAGAGCCTAAACATCC
KIT Exon9 301-400 bases S eq. .
Primer S equence ID 2941 Left TTTGTTTTAAAAGTATGCCACATCC
2921 Left CTCACTAGGTCACCAAAGTGCTTAT 2942 Right TATGGTAGACAGAGCCTAAACATCC
2922 Right CCCCTTAAATTGGATTAAAAAGAAA
2943 Left CTCACTAGGTCACCAAAGTGCTTAT
2923 Left CTCACTAGGTCACCAAAGTGCTTAT 2944 Right ATCCCCTTAAATTGGATTAAAAAGA
2924 Right AACATCCCCTTAAATTGGATTAAAA
__________________________________________ 2945 Left CCACATCCCAAGTGTTTTATGTATT
2925 Left CCACATCCCAAGTGTTTTATGTATT 2946 Right TCATGACTGATATGGTAGACAGAGC
2926 Right TATGGTAGACAGAGCCTAAACATCC _____ 2947 Left CACTAGGTCACCAAAGTGCTTATTC
2927 Left CTCACTAGGTCACCAAAGTGCTTAT 2948 Right AACATCCCCTTAAATTGGATTAAAA
2928 Right CATCCCCTTAAATTGGATTAAAAAG ____ 2949 Left TCACTAGGTCACCAAAGTGCTTATT
2929 Left CCACATCCCAAGTGTTTTATGTATT 2950 Right AACATCCCCTTAAATTGGATTAAAA
2930 Right GTGATGCATGTATTACCAGAAATGA
2951 Left TTTGTTTTAAAAGTATGCCACATCC
2931 Left CCACATCCCAAGTGTTTTATGTATT 2952 Right GTGATGCATGTATTACCAGAAATGA
2932 Right AACATCCCCTTAAATTGGATTAAAA
S eq. . 2953 Left TCACCAAAGTGCTTATTCTTAGACA
Pnmer S equence ID 2954 Right CCCCTTAAATTGGATTAAAAAGAAA
2933 Left CACTAGGTCACCAAAGTGCTTATTC ______ 2934 Right CCCCTTAAATTGGATTAAAAAGAAA 2955 Left ACTCACTAGGTCACCAAAGTGCTTA
2956 Right CCCCTTAAATTGGATTAAAAAGAAA
2935 Left TCACTAGGTCACCAAAGTGCTTATT ______ 2936 Right CCCCTTAAATTGGATTAAAAAGAAA 2957 Left TTTGTTTTAAAAGTATGCCACATCC
2958 Right AACATCCCCTTAAATTGGATTAAAA
2937 Left CTCACTAGGTCACCAAAGTGCTTAT ______ 2938 Right TCCCCTTAAATTGGATTAAAAAGAA 2959 Left AGGTCACCAAAGTGCTTATTCTTAG
2960 Right CCCCTTAAATTGGATTAAAAAGAAA
2939 Left TTTGTTTTAAAAGTATGCCACATCC
2940 Right CCCCTTAAATTGGATTAAAAAGAAA
KIT Exon9 401-500 bases S eq. 2962 Right TATGGTAGACAGAGCCTAAACATCC
ID Primer Sequence 2961 Left CTCACTAGGTCACCAAAGTGCTTAT 2963 Left TACAGTCGTAGAAACTCAGTGTTGG

2964 Right CCCCTTAAATTGGATTAAAAAGAAA 2982 Right GTGATGCATGTATTACCAGAAATGA
2965 Left CTCACTAGGTCACCAAAGTGCTTAT 2983 Left TCACTAGGTCACCAAAGTGCTTATT
2966 Right GTGATGCATGTATTACCAGAAATGA 2984 Right GTGATGCATGTATTACCAGAAATGA
2967 Left CTCACTAGGTCACCAAAGTGCTTAT 2985 Left TACAGTCGTAGAAACTCAGTGTTGG
2968 Right TCATGACTGATATGGTAGACAGAGC 2986 Right ATCCCCTTAAATTGGATTAAAAAGA
2969 Left TACAGTCGTAGAAACTCAGTGTTGG 2987 Left TCACCAAAGTGCTTATTCTTAGACA
2970 Right CATCCCCTTAAATTGGATTAAAAAG 2988 Right TATGGTAGACAGAGCCTAAACATCC
2971 Left CACTAGGTCACCAAAGTGCTTATTC 2989 Left ACTCACTAGGTCACCAAAGTGCTTA
2972 Right TATGGTAGACAGAGCCTAAACATCC 2990 Right TATGGTAGACAGAGCCTAAACATCC
2973 Left TCACTAGGTCACCAAAGTGCTTATT 2991 Left ACAGTCGTAGAAACTCAGTGTTGGT
2974 Right TATGGTAGACAGAGCCTAAACATCC 2992 Right AACATCCCCTTAAATTGGATTAAAA
2975 Left ACAGTCGTAGAAACTCAGTGTTGGT 2993 Left CTAGGTCACCAAAGTGCTTATTCTT
2976 Right CCCCTTAAATTGGATTAAAAAGAAA 2994 Right TATGGTAGACAGAGCCTAAACATCC
2977 Left TACAGTCGTAGAAACTCAGTGTTGG 2995 Left AGGTCACCAAAGTGCTTATTCTTAG
2978 Right TCCCCTTAAATTGGATTAAAAAGAA 2996 Right TATGGTAGACAGAGCCTAAACATCC
2979 Left CTCACTAGGTCACCAAAGTGCTTAT 2997 Left TCACCAAAGTGCTTATTCTTAGACA
2980 Right GGTCAATGTTGGAATGAACTTAAAA 2998 Right GTGATGCATGTATTACCAGAAATGA
S eq. . 2999 Left CACTAGGTCACCAAAGTGCTTATTC

Pnmer S equence ID 3000 Right TCATGACTGATATGGTAGACAGAGC
2981 Left CACTAGGTCACCAAAGTGCTTATTC
KIT Exon9 501-600 bases S eq. .
Primer S equence ID 3015 Left TCCTCCTCTATGCTATTTCTTTTCA

3001 Left CCTCTATGCTATTTCTTTTCAACCA 3016 Right TATGGTAGACAGAGCCTAAACATCC
3002 Right TATGGTAGACAGAGCCTAAACATCC
3017 Left ATTTTATTGAATTCCTTTCCAATCC
3003 Left CTCACTAGGTCACCAAAGTGCTTAT 3018 Right GTGATGCATGTATTACCAGAAATGA
3004 Right AGGCAGTGTTAACTTTTGGATACAG
S eq. . 3019 Left CTCACTAGGTCACCAAAGTGCTTAT

Pnmer S equence ID 3020 Right GTAAATATATTCCCCCATTTGCTTT
3005 Left TTTTATGCTTTCCTCCTCTATGCTA _____ 3006 Right CCCCTTAAATTGGATTAAAAAGAAA 3021 Left GCTTTCCTCCTCTATGCTATTTCTT
3022 Right AACATCCCCTTAAATTGGATTAAAA
3007 Left CTCACTAGGTCACCAAAGTGCTTAT _____ 3008 Right GGCAGTGTTAACTTTTGGATACAGT 3023 Left TTTTATGCTTTCCTCCTCTATGCTA
3024 Right AACATCCCCTTAAATTGGATTAAAA
3009 Left GCTTTCCTCCTCTATGCTATTTCTT
3010 Right TATGGTAGACAGAGCCTAAACATCC 3025 Left CCACATCCCAAGTGTTTTATGTATT
3026 Right GGCAGTGTTAACTTTTGGATACAGT
3011 Left TTTTATGCTTTCCTCCTCTATGCTA _____ 3012 Right TATGGTAGACAGAGCCTAAACATCC 3027 Left TTTAGTAGAGACGAGGTTTCACCAT
3028 Right CCCCTTAAATTGGATTAAAAAGAAA
3013 Left CCACATCCCAAGTGTTTTATGTATT _____ 3014 Right AGGCAGTGTTAACTTTTGGATACAG 3029 Left GCTGAGATTACAGGTGTGAGCTACT

3030 Right CCCCTTAAATTGGATTAAAAAGAAA 035 Left GCTGAGATTACAGGTGTGAGCTACT
3 036 Right TATGGTAGACAGAGCCTAAACATCC
3031 Left CTCACTAGGTCACCAAAGTGCTTAT
3032 Right GATTGTTCTAATTCTGTTTGGGTGT 3 037 Left CACTAGGTCACCAAAGTGCTTATTC
3 038 Right AGGCAGTGTTAACTTTTGGATACAG
3033 Left TACAGTCGTAGAAACTCAGTGTTGG
3034 Right GTAAATATATTCCCCCATTTGCTTT 3 039 Left TCACTAGGTCACCAAAGTGCTTATT
3 040 Right AGGCAGTGTTAACTTTTGGATACAG
KIT Exon9 801-1000 bases S eq. .
Primer S equence ID 2061 Left CCCTGTTTTACAGTCGTAGAAACTC
3041 Left ATTCCTTTCCAATCCTTTCAGTAAC 2062 Right AGGCAGTGTTAACTTTTGGATACAG
3042 Right AGGCAGTGTTAACTTTTGGATACAG
2063 Left ATTCCTTTCCAATCCTTTCAGTAAC
3043 Left ATTTTATTGAATTCCTTTCCAATCC 2064 Right GTAAATATATTCCCCCATTTGCTTT
3044 Right AGGCAGTGTTAACTTTTGGATACAG
2065 Left TACATCCTTGATTTTGTTGTTGTTG
3045 Left TACAGTCGTAGAAACTCAGTGTTGG 2066 Right AACATCCCCTTAAATTGGATTAAAA
3046 Right AGGCAGTGTTAACTTTTGGATACAG
2067 Left AACCCTCTGCAATGGGTATTACTAT
3047 Left TACATCCTTGATTTTGTTGTTGTTG 2068 Right AGGCAGTGTTAACTTTTGGATACAG
3048 Right CCCCTTAAATTGGATTAAAAAGAAA
2069 Left TTATTGAATTCCTTTCCAATCCTTT
3049 Left ATTCCTTTCCAATCCTTTCAGTAAC 2070 Right AGGCAGTGTTAACTTTTGGATACAG
3050 Right GGCAGTGTTAACTTTTGGATACAGT
2071 Left TTTTATTGAATTCCTTTCCAATCCT
3051 Left CCTCTATGCTATTTCTTTTCAACCA 2072 Right AGGCAGTGTTAACTTTTGGATACAG
3052 Right GTAAATATATTCCCCCATTTGCTTT
__________________________________________ 2073 Left TTTATTGAATTCCTTTCCAATCCTT
3053 Left ATTTTATTGAATTCCTTTCCAATCC 2074 Right AGGCAGTGTTAACTTTTGGATACAG
3054 Right GGCAGTGTTAACTTTTGGATACAGT _____ 2075 Left ATTTTATTGAATTCCTTTCCAATCC
3055 Left TTCTGGTCTACATCCTTGATTTTGT 2076 Right GTAAATATATTCCCCCATTTGCTTT
3056 Right CCCCTTAAATTGGATTAAAAAGAAA
2077 Left CCTCTATGCTATTTCTTTTCAACCA
3057 Left TACAGTCGTAGAAACTCAGTGTTGG 2078 Right TGCTTTCTCTAGCTCTTTTTAATGG
3058 Right GGCAGTGTTAACTTTTGGATACAGT
S eq. . 2079 Left CCACATCCCAAGTGTTTTATGTATT
Pnmer S equence ID 2080 Right ACTACTCAAAACCTGAGAAAACACG
3059 Left CCTCTATGCTATTTCTTTTCAACCA
3060 Right GATTGTTCTAATTCTGTTTGGGTGT
KIT Exon9 2kb S eq. .
Primer S equence ID 2087 Left TGTAGGTGTTAAGAAAGGAAACAGG
3081 Left TGTAGGTGTTAAGAAAGGAAACAGG 2088 Right ACTACTCAAAACCTGAGAAAACACG
3082 Right GCTTCCTTTATGGACGGTTTATATT
__________________________________________ 2089 Left AAGTAATGATCGCACAATTACAACA
3083 Left TCCTCACTTTGACTATTTTCTGGTC 2090 Right CCCCTTAAATTGGATTAAAAAGAAA
3084 Right GCTTCCTTTATGGACGGTTTATATT _____ 2091 Left CCAAGTTACCAAAAAGTAATGATCG
3085 Left GTTAAGAAAGGAAACAGGGAATAGC 2092 Right CCCCTTAAATTGGATTAAAAAGAAA
3086 Right GCTTCCTTTATGGACGGTTTATATT

3093 Left TGGATAAGCTTGTTCTAGTGGGTAG
3094 Right ACTACTCAAAACCTGAGAAAACACG 3 107 Left GGTCTACATCCTTGATTTTGTTGTT
3 108 Right GCTTCCTTTATGGACGGTTTATATT
3095 Left TGTAGGTGTTAAGAAAGGAAACAGG
3096 Right CCTCACTACTCAAAACCTGAGAAAA 3 109 Left GGAATAAGCCTCTTTATCACAACAA
3 110 Right GTAAATATATTCCCCCATTTGCTTT
3097 Left ATAACTAGGCCTTCCTGCTTAGAAC
3098 Right GCTTCCTTTATGGACGGTTTATATT 2111 Left TGTAGGTGTTAAGAAAGGAAACAGG
3 112 Right TCTTTAAGCTTTCCTGTATTTTCCA
3099 Left GTTAAGAAAGGAAACAGGGAATAGC
3100 Right CCTCACTACTCAAAACCTGAGAAAA 3 113 Left AAGTAATGATCGCACAATTACAACA
3 114 Right AACATCCCCTTAAATTGGATTAAAA
Seq. .
Pnmer S equence ID 3 115 Left TAACTAGGCCTTCCTGCTTAGAACT
3101 Left TGGATAAGCTTGTTCTAGTGGGTAG 3 116 Right GCTTCCTTTATGGACGGTTTATATT
3102 Right CCTCACTACTCAAAACCTGAGAAAA
3 117 Left TTCATGGAATAAGCCTCTTTATCAC
3103 Left TCTGGTCTACATCCTTGATTTTGTT 3 118 Right GTAAATATATTCCCCCATTTGCTTT
3104 Right GCTTCCTTTATGGACGGTTTATATT
3 119 Left TTCTTTTGGGGAATACTAAGTAGGG
3105 Left TTCTGGTCTACATCCTTGATTTTGT 3 120 Right GTAAATATATTCCCCCATTTGCTTT
3106 Right GCTTCCTTTATGGACGGTTTATATT
KIT Exon9-10 2kb Seq. . 3 139 Left AACCCTCTGCAATGGGTATTACTAT
Pnmer Sequence ID 1' 140 Right ATTAGAGCACTCTGGAGAGAGAACA
3121 Left CCTCTATGCTATTTCTTTTCAACCA
3122 Right ATTAGAGCACTCTGGAGAGAGAACA 1; 141 Left GCTGAGATTACAGGTGTGAGCTACT
1' 142 Right ATTAGAGCACTCTGGAGAGAGAACA
3123 Left ATTCCTTTCCAATCCTTTCAGTAAC
3124 Right AGCACTCTGGAGAGAGAACAAATAA 3 143 Left ATTTTATTGAATTCCTTTCCAATCC
__________________________________________ 3 144 Right CTCTGGAGAGAGAACAAATAAATGG
3125 Left ATTCCTTTCCAATCCTTTCAGTAAC
3126 Right ATTAGAGCACTCTGGAGAGAGAACA 145 Left TTATTGAATTCCTTTCCAATCCTTT
1' 146 Right ATTAGAGCACTCTGGAGAGAGAACA
3127 Left ATTTTATTGAATTCCTTTCCAATCC
3128 Right ATTAGAGCACTCTGGAGAGAGAACA 3 147 Left TTTATTGAATTCCTTTCCAATCCTT
1' 148 Right ATTAGAGCACTCTGGAGAGAGAACA
3129 Left GCTTTCCTCCTCTATGCTATTTCTT
3130 Right ATTAGAGCACTCTGGAGAGAGAACA 1; 149 Left TTTTATTGAATTCCTTTCCAATCCT
Seq. . 1; 150 Right ATTAGAGCACTCTGGAGAGAGAACA
ID Pnmer Sequence 3131 Left AACCCTCTGCAATGGGTATTACTAT 1; 151 Left ATGCTTTCCTCCTCTATGCTATTTC
3132 Right AGCACTCTGGAGAGAGAACAAATAA 3 152 Right AGCACTCTGGAGAGAGAACAAATAA
3133 Left GCTGAGATTACAGGTGTGAGCTACT 1; 153 Left TTTTATGCTTTCCTCCTCTATGCTA
3134 Right AGCACTCTGGAGAGAGAACAAATAA 3 154 Right CTCTGGAGAGAGAACAAATAAATGG
3135 Left ATTCCTTTCCAATCCTTTCAGTAAC 3 155 Left ATGCTTTCCTCCTCTATGCTATTTC
3136 Right GCACTCTGGAGAGAGAACAAATAAA 3 156 Right ATTAGAGCACTCTGGAGAGAGAACA
3137 Left ATTCCTTTCCAATCCTTTCAGTAAC 3 157 Left ATTCCTTTCCAATCCTTTCAGTAAC
3138 Right CTCTGGAGAGAGAACAAATAAATGG 3 158 Right GAGCACTCTGGAGAGAGAACAAATA

3159 Left ATTCCTTTCCAATCCTTTCAGTAAC 3 160 Right TCTGGAGAGAGAACAAATAAATGGT
KIT Exon9-11 2kb Seq. Seq.
Primer Sequence Primer Sequence ID ID
3161 Left CCACATCCCAAGTGTTTTATGTATT 3 181 Left TCACCAAAGTGCTTATTCTTAGACA
3162 Right TTCTCTATGGCAAACCTATCAAAAG 3 182 Right GTTCTCTATGGCAAACCTATCAAAA
3163 Left CCACATCCCAAGTGTTTTATGTATT 3 183 Left CTAGGTCACCAAAGTGCTTATTCTT
3164 Right GTTCTCTATGGCAAACCTATCAAAA 3 184 Right TTCTCTATGGCAAACCTATCAAAAG
3165 Left CCACATCCCAAGTGTTTTATGTATT 2185 Left TTTGTTTTAAAAGTATGCCACATCC
3166 Right ATGTTGTCCAGAGACATTTTCCTAC 3 186 Right CATTTTCCTACGATGTTCTCTATGG
3167 Left CACTAGGTCACCAAAGTGCTTATTC 3 187 Left CCACATCCCAAGTGTTTTATGTATT
3168 Right TTCTCTATGGCAAACCTATCAAAAG 3 188 Right GGAATTAAAAACAATGTTGTCCAGA
3169 Left CCACATCCCAAGTGTTTTATGTATT 3 189 Left TTTGTTTTAAAAGTATGCCACATCC
3170 Right CATTTTCCTACGATGTTCTCTATGG 3 190 Right AATGTTGTCCAGAGACATTTTCCTA
3171 Left CCACATCCCAAGTGTTTTATGTATT 3 191 Left CTAGGTCACCAAAGTGCTTATTCTT
3172 Right ATGTTCTCTATGGCAAACCTATCAA 3 192 Right GTTCTCTATGGCAAACCTATCAAAA
3173 Left CCACATCCCAAGTGTTTTATGTATT 3 193 Left AGGTCACCAAAGTGCTTATTCTTAG
3174 Right AATGTTGTCCAGAGACATTTTCCTA 3 194 Right GTTCTCTATGGCAAACCTATCAAAA
3175 Left CCACATCCCAAGTGTTTTATGTATT 2195 Left TTTGTTTTAAAAGTATGCCACATCC
3176 Right AGGAATTAAAAACAATGTTGTCCAG 3 196 Right AGGAATTAAAAACAATGTTGTCCAG
3177 Left TCACCAAAGTGCTTATTCTTAGACA 3 197 Left TTTGTTTTAAAAGTATGCCACATCC
3178 Right TTCTCTATGGCAAACCTATCAAAAG 3 198 Right TTGTGCAGTTTCAAAATCAATAAAG
3179 Left TTTGTTTTAAAAGTATGCCACATCC 3 199 Left TCACCAAAGTGCTTATTCTTAGACA
3180 Right ATGTTGTCCAGAGACATTTTCCTAC 3200 Right ATGTTCTCTATGGCAAACCTATCAA
KIT Exon10 130-150 bases Seq. . 2211 Left CACATTTCTCTTCCATTGTA
Primer Sequence ID 2 212 Right GAGAGAACAAATAAATGGTTAC
3201 Left TCCACATTTCTCTTCCATTGTA
3202 Right GAGAACAAATAAATGGTTAC 2213 Left CCACATTTCTCTTCCATTGTA
Seq. . 2 214 Right AGAGAACAAATAAATGGTTA
ID Primer Sequence 3203 Left CCACATTTCTCTTCCATTGTA 2215 Left CCACATTTCTCTTCCATTGTA
3204 Right AGAGAACAAATAAATGGTTAC 2 216 Right GAGAACAAATAAATGGTTAC
3205 Left TCCACATTTCTCTTCCATTGT
3206 Right GAGAACAAATAAATGGTTAC 2217 Left TCCACATTTCTCTTCCATTG
2 218 Right GAGAACAAATAAATGGTTAC
3207 Left CCACATTTCTCTTCCATTGT
3208 Right AGAGAACAAATAAATGGTTAC 2219 Left TCCACATTTCTCTTCCATTGT
2 220 Right GAGAACAAATAAATGGTTA
3209 Left TCCACATTTCTCTTCCATTGTA
3210 Right GAGAACAAATAAATGGTTA 2221 Left CCACATTTCTCTTCCATTG
2 222 Right AGAGAACAAATAAATGGTTAC

3223 Left CACATTTCTCTTCCATTGT 2233 Left CACATTTCTCTTCCATTGTA
3224 Right GAGAGAACAAATAAATGGTTAC 2234 Right GAGAGAACAAATAAATGGTTA
3225 Left CCACATTTCTCTTCCATTGT 2235 Left CCACATTTCTCTTCCATTGTA
3226 Right AGAGAACAAATAAATGGTTA 2236 Right GAGAACAAATAAATGGTTA
3227 Left CCACATTTCTCTTCCATTGT 2237 Left CACATTTCTCTTCCATTGTA
3228 Right GAGAACAAATAAATGGTTAC 2238 Right AGAGAACAAATAAATGGTTAC
3229 Left CATTTCTCTTCCATTGTA 2239 Left TCCACATTTCTCTTCCATTG
3230 Right GAGAGAGAACAAATAAATGGTTAC 2240 Right GAGAACAAATAAATGGTTA
3231 Left ACATTTCTCTTCCATTGTA
3232 Right AGAGAGAACAAATAAATGGTTAC
KIT Exon10 151-200 bases Seq. .
Pnmer Sequence ID 3261 Left AAAGTTTGTGATTCCACATTTCTCT

3241 Left AGTTTGTGATTCCACATTTCTCTTC 3262 Right GAGCACTCTGGAGAGAGAACAAATA
3242 Right AGCACTCTGGAGAGAGAACAAATAA
3263 Left CAAAGTTTGTGATTCCACATTTCTC
3243 Left AGTTTGTGATTCCACATTTCTCTTC 3264 Right AGCACTCTGGAGAGAGAACAAATAA
3244 Right GCACTCTGGAGAGAGAACAAATAAA ____ 3265 Left CAAAGTTTGTGATTCCACATTTCTC
3245 Left AGTTTGTGATTCCACATTTCTCTTC 3266 Right GCACTCTGGAGAGAGAACAAATAAA
3246 Right GAGCACTCTGGAGAGAGAACAAATA
3267 Left CAAAGTTTGTGATTCCACATTTCTC
3247 Left AGTTTGTGATTCCACATTTCTCTTC 3268 Right CTCTGGAGAGAGAACAAATAAATGG
3248 Right TCTGGAGAGAGAACAAATAAATGGT
3269 Left AGTTTGTGATTCCACATTTCTCTTC
3249 Left AAGTTTGTGATTCCACATTTCTCTT 3270 Right CTCTGGAGAGAGAACAAATAAATGGT
3250 Right AGCACTCTGGAGAGAGAACAAATAA
3271 Left CAAAGTTTGTGATTCCACATTTCTC
3251 Left AAAGTTTGTGATTCCACATTTCTCT 3272 Right GAGCACTCTGGAGAGAGAACAAATA
3252 Right AGCACTCTGGAGAGAGAACAAATAA
3273 Left CAAAGTTTGTGATTCCACATTTCTC
3253 Left GATTCCACATTTCTCTTCCATTGTA 3274 Right TCTGGAGAGAGAACAAATAAATGGT
3254 Right AGCACTCTGGAGAGAGAACAAATAA
3275 Left CAAAGTTTGTGATTCCACATTTCT
3255 Left AAGTTTGTGATTCCACATTTCTCTT 3276 Right AGCACTCTGGAGAGAGAACAAATAA
3256 Right GCACTCTGGAGAGAGAACAAATAAA ____ S eq. . 3277 Left GTGATTCCACATTTCTCTTCCATT
Pnmer Sequence ID 3278 Right AGCACTCTGGAGAGAGAACAAATAA
3257 Left AAAGTTTGTGATTCCACATTTCTCT _____ 3258 Right GCACTCTGGAGAGAGAACAAATAAA 3279 Left CAAAGTTTGTGATTCCACATTTCT
3280 Right ATTAGAGCACTCTGGAGAGAGAACA
3259 Left AAGTTTGTGATTCCACATTTCTCTT
3260 Right GAGCACTCTGGAGAGAGAACAAATA
KIT Exon10 201-300 bases S eq. 3282 Right AGCACTCTGGAGAGAGAACAAATAA
ID Primer Sequence 3281 Left GTACAATGTAACCAAGGTGAAGCTC 3283 Left GAGTACAATGTAACCAAGGTGAAGC

3284 Right AGCACTCTGGAGAGAGAACAAATAA 3302 Right GAGCACTCTGGAGAGAGAACAAATA
3285 Left TACAATGTAACCAAGGTGAAGCTCT 3303 Left GAGTACAATGTAACCAAGGTGAAGC
3286 Right AGCACTCTGGAGAGAGAACAAATAA 3304 Right GAGCACTCTGGAGAGAGAACAAATA
3287 Left TACAATGTAACCAAGGTGAAGCTCT 3305 Left GTACAATGTAACCAAGGTGAAGCTC
3288 Right ATTAGAGCACTCTGGAGAGAGAACA 3306 Right TCTGGAGAGAGAACAAATAAATGGT
3289 Left GTACAATGTAACCAAGGTGAAGCTC 3307 Left GAGTACAATGTAACCAAGGTGAAGC
3290 Right GCACTCTGGAGAGAGAACAAATAAA 3308 Right TCTGGAGAGAGAACAAATAAATGGT
3291 Left GAGTACAATGTAACCAAGGTGAAGC 3309 Left TACAATGTAACCAAGGTGAAGCTCT
3292 Right GCACTCTGGAGAGAGAACAAATAAA 3310 Right GAGCACTCTGGAGAGAGAACAAATA
3293 Left GTACAATGTAACCAAGGTGAAGCTC 3311 Left TACAATGTAACCAAGGTGAAGCTCT
3294 Right CTCTGGAGAGAGAACAAATAAATGG 3312 Right TCTGGAGAGAGAACAAATAAATGGT
3295 Left GAGTACAATGTAACCAAGGTGAAGC 3313 Left CTCTGAGACTCACATAGCTTTGCAT
3296 Right CTCTGGAGAGAGAACAAATAAATGG 3314 Right AGCACTCTGGAGAGAGAACAAATAA
3297 Left TACAATGTAACCAAGGTGAAGCTCT 3315 Left GTACAATGTAACCAAGGTGAAGCTC
3298 Right GCACTCTGGAGAGAGAACAAATAAA 3316 Right TAGAGCACTCTGGAGAGAGAACAAA
3299 Left TACAATGTAACCAAGGTGAAGCTCT 3317 Left CTCTGAGACTCACATAGCTTTGCAT
3300 Right CTCTGGAGAGAGAACAAATAAATGG 3318 Right ATTAGAGCACTCTGGAGAGAGAACA
S eq. . 3319 Left TACAATGTAACCAAGGTGAAGCTC
Primer Sequence ID 3320 Right AGCACTCTGGAGAGAGAACAAATAA
3301 Left GTACAATGTAACCAAGGTGAAGCTC
KIT Exon10 301-400 bases S eq. .
Primer S equence ID 3335 Left TCTGCAGTATTGTGGTTTCAAGTTA

3321 Left TCTATTCTGCAGTATTGTGGTTTCA 3336 Right GCACTCTGGAGAGAGAACAAATAAA
3322 Right AGCACTCTGGAGAGAGAACAAATAA
3337 Left TATTCTGCAGTATTGTGGTTTCAAG
3323 Left TCTGCAGTATTGTGGTTTCAAGTTA 3338 Right AGCACTCTGGAGAGAGAACAAATAA
3324 Right AGCACTCTGGAGAGAGAACAAATAA
3339 Left CTATTCTGCAGTATTGTGGTTTCAA
3325 Left TCTATTCTGCAGTATTGTGGTTTCA 1' 340 Right AGCACTCTGGAGAGAGAACAAATAA
3326 Right ATTAGAGCACTCTGGAGAGAGAACA
1' 341 Left TCTGCAGTATTGTGGTTTCAAGTTA
3327 Left TCTGCAGTATTGTGGTTTCAAGTTA 1' 342 Right CTCTGGAGAGAGAACAAATAAATGG
3328 Right ATTAGAGCACTCTGGAGAGAGAACA
S eq. . 1' 343 Left GAGTACAATGTAACCAAGGTGAAGC
Pnmer S equence ID 3344 Right ATTAGAGCACTCTGGAGAGAGAACA
3329 Left TCTATTCTGCAGTATTGTGGTTTCA _____ 3330 Right GCACTCTGGAGAGAGAACAAATAAA 3345 Left GTACAATGTAACCAAGGTGAAGCTC
1' 346 Right ATTAGAGCACTCTGGAGAGAGAACA
3331 Left ATTCTGCAGTATTGTGGTTTCAAGT _____ 3332 Right AGCACTCTGGAGAGAGAACAAATAA 3347 Left ATTCTGCAGTATTGTGGTTTCAAGT
1' 348 Right ATTAGAGCACTCTGGAGAGAGAACA
3333 Left TCTATTCTGCAGTATTGTGGTTTCA _____ 3334 Right CTCTGGAGAGAGAACAAATAAATGG

3349 Left TATTCTGCAGTATTGTGGTTTCAAG 3355 Left TCTGCAGTATTGTGGTTTCAAGTTA
3350 Right ATTAGAGCACTCTGGAGAGAGAACA 3356 Right GAGCACTCTGGAGAGAGAACAAATA
3351 Left CTATTCTGCAGTATTGTGGTTTCAA 3357 Left TCTGCAGTATTGTGGTTTCAAGTTA
3352 Right ATTAGAGCACTCTGGAGAGAGAACA 3358 Right TCTGGAGAGAGAACAAATAAATGGT
3353 Left TCTATTCTGCAGTATTGTGGTTTCA 3359 Left ATTCTGCAGTATTGTGGTTTCAAGT
3354 Right TCTGGAGAGAGAACAAATAAATGGT 3360 Right GCACTCTGGAGAGAGAACAAATAAA
KIT Exonll 151-200 bases Seq.
Primer Sequence Primer Sequence ID ID
3361 Left AAGGTGATCTATTTTTCCCTTTCTC 2381 Left GGTGATCTATTTTTCCCTTTCTCC
3362 Right GAAAGCCCCTGTTTCATACTGAC 2382 Right AAAGCCCCTGTTTCATACTGAC
3363 Left AAGGTGATCTATTTTTCCCTTTCTC 2383 Left AAAGGTGATCTATTTTTCCCTTTCTC
3364 Right AAAGCCCCTGTTTCATACTGAC 2384 Right AAAGCCCCTGTTTCATACTGAC
3365 Left AAAGGTGATCTATTTTTCCCTTTCT 2385 Left CTATTTTTCCCTTTCTCCCCACAG
3366 Right GAAAGCCCCTGTTTCATACTGAC 2386 Right TTATGTGTACCCAAAAAGGTGACAT
3367 Left AAAGGTGATCTATTTTTCCCTTTCT 2387 Left AAGGTGATCTATTTTTCCCTTTCTC
3368 Right AAAGCCCCTGTTTCATACTGAC 2388 Right GGAAAGCCCCTGTTTCATACT
3369 Left AGGTGATCTATTTTTCCCTTTCTCC 2389 Left AAAGGTGATCTATTTTTCCCTTTCT
3370 Right GAAAGCCCCTGTTTCATACTGAC 2390 Right ATGGAAAGCCCCTGTTTCATACT
3371 Left AAGGTGATCTATTTTTCCCTTTCTC 2391 Left AAGGTGATCTATTTTTCCCTTTCTC
3372 Right ATGGAAAGCCCCTGTTTCATACT 2392 Right ATGGAAAGCCCCTGTTTCATA
3373 Left GGTGATCTATTTTTCCCTTTCTCC 2393 Left TATTTTTCCCTTTCTCCCCACAG
3374 Right GAAAGCCCCTGTTTCATACTGAC 2394 Right TTATGTGTACCCAAAAAGGTGACAT
3375 Left AAAGGTGATCTATTTTTCCCTTTCTC 2395 Left CTATTTTTCCCTTTCTCCCCACA
3376 Right GAAAGCCCCTGTTTCATACTGAC 2396 Right TTATGTGTACCCAAAAAGGTGACAT
3377 Left AGGTGATCTATTTTTCCCTTTCTCC 2397 Left AAAGGTGATCTATTTTTCCCTTTCT
3378 Right AAAGCCCCTGTTTCATACTGAC 2398 Right CAAAAAGGTGACATGGAAAGC
3379 Left AAGGTGATCTATTTTTCCCTTTCTC 2399 Left AAAGGTGATCTATTTTTCCCTTTCT
3380 Right ATGGAAAGCCCCTGTTTCATAC 2,400 Right ATGGAAAGCCCCTGTTTCATAC
KIT Exonll 201-300 bases S eq. . 3405 Left GTTCTCTCTCCAGAGTGCTCTAATG
Pnmer Sequence ID 3406 Right TTATGTGTACCCAAAAAGGTGACAT
3401 Left TTATTTGTTCTCTCTCCAGAGTGCT
3402 Right TTATGTGTACCCAAAAAGGTGACAT 3407 Left CAGAGTGCTCTAATGACTGAGACAA
3408 Right TTATGTGTACCCAAAAAGGTGACAT
3403 Left TGTTCTCTCTCCAGAGTGCTCTAAT
3404 Right TTATGTGTACCCAAAAAGGTGACAT 3409 Left CTCTCTCCAGAGTGCTCTAATGACT
3410 Right TTATGTGTACCCAAAAAGGTGACAT

3425 Left GTTCTCTCTCCAGAGTGCTCTAATG
3411 Le ft TTATTTGTTCTCTCTCCAGAGTGCT 3426 Right TGTTATGTGTACCCAAAAAGGTGAC
3412 Right TGTTATGTGTACCCAAAAAGGTGAC
3427 Left TGTTCTCTCTCCAGAGTGCTCTAAT
3413 Le ft TTATTTGTTCTCTCTCCAGAGTGCT 3428 Right GTTATGTGTACCCAAAAAGGTGACA
3414 Right GTTATGTGTACCCAAAAAGGTGACA
3429 Left GTTCTCTCTCCAGAGTGCTCTAATG
3415 Le ft AAGGTGATCTATTTTTCCCTTTCTC 3430 Right GTTATGTGTACCCAAAAAGGTGACA
3416 Right TTATGTGTACCCAAAAAGGTGACAT
3431 Left GTTCTCTCTCCAGAGTGCTCTAATG
3417 Le ft AAGGTGATCTATTTTTCCCTTTCTC 3432 Right CTGTTATGTGTACCCAAAAAGGTG
3418 Right GCAATTTCACAGAAAACTCATTGTT
3433 Left TGTTCTCTCTCCAGAGTGCTCTAAT
3419 Le ft TTATTTGTTCTCTCTCCAGAGTGCT 3434 Right CTGTTATGTGTACCCAAAAAGGTG
3420 Right CTGTTATGTGTACCCAAAAAGGTG
3435 Left CAGAGTGCTCTAATGACTGAGACAA
Seq. . 3436 Right GTTATGTGTACCCAAAAAGGTGACA
ID Pnmer Sequence 3421 Le ft TCTCTCTCCAGAGTGCTCTAATGAC 3437 Left CAGAGTGCTCTAATGACTGAGACAA
3422 Right TTATGTGTACCCAAAAAGGTGACAT 3438 Right TGTTATGTGTACCCAAAAAGGTGAC
3423 Le ft TGTTCTCTCTCCAGAGTGCTCTAAT 3439 Left TTATTTGTTCTCTCTCCAGAGTGCT
3424 Right TGTTATGTGTACCCAAAAAGGTGAC 3440 Right CTGTTATGTGTACCCAAAAAGGTGA
KIT Exonll 301-400 bases Seq. . 2455 Left CAGAGTGCTCTAATGACTGAGACAA

Pnmer Sequence ID 2456 Right GTTCTCTATGGCAAACCTATCAAAA
3441 Le ft TTATTTGTTCTCTCTCCAGAGTGCT
3442 Right TTCTCTATGGCAAACCTATCAAAAG 2457 Left TTATTTGTTCTCTCTCCAGAGTGCT
2458 Right ATGTTGTCCAGAGACATTTTCCTAC
3443 Le ft TTATTTGTTCTCTCTCCAGAGTGCT
3444 Right GTTCTCTATGGCAAACCTATCAAAA 2459 Le ft TGTTCTCTCTCCAGAGTGCTCTAAT
2460 Right ATGTTGTCCAGAGACATTTTCCTAC
3445 Le ft TGTTCTCTCTCCAGAGTGCTCTAAT
3446 Right TTCTCTATGGCAAACCTATCAAAAG
3447 Le ft GTTCTCTCTCCAGAGTGCTCTAATG 2461 Le ft GTTCTCTCTCCAGAGTGCTCTAATG
3448 Right TTCTCTATGGCAAACCTATCAAAAG 2462 Right ATGTTGTCCAGAGACATTTTCCTAC
3449 Left CAGAGTGCTCTAATGACTGAGACAA 2463 Left TTATTTGTTCTCTCTCCAGAGTGCT
3450 Right TTCTCTATGGCAAACCTATCAAAAG 2464 Right CATTTTCCTACGATGTTCTCTATGG
3465 Le ft CTCTCTCCAGAGTGCTCTAATGACT
3466 Right TTCTCTATGGCAAACCTATCAAAAG
Seq. .
Primer Sequence ID 2467 Left TTATTTGTTCTCTCTCCAGAGTGCT

3451 Left GTTCTCTCTCCAGAGTGCTCTAATG 2468 Right ATGTTCTCTATGGCAAACCTATCAA
3452 Right GTTCTCTATGGCAAACCTATCAAAA ____ 2469 Left CAGAGTGCTCTAATGACTGAGACAA
3453 Left TGTTCTCTCTCCAGAGTGCTCTAAT 2470 Right ATGTTGTCCAGAGACATTTTCCTAC
3454 Right GTTCTCTATGGCAAACCTATCAAAA ____ 2471 Left TTATTTGTTCTCTCTCCAGAGTGCT

3472 Right AATGTTGTCCAGAGACATTTTCCTA
2477 Left TGTTCTCTCTCCAGAGTGCTCTAAT
3473 Left CTCTCTCCAGAGTGCTCTAATGACT 2478 Right CATTTTCCTACGATGTTCTCTATGG
3474 Right GTTCTCTATGGCAAACCTATCAAAA
2479 Left GTTCTCTCTCCAGAGTGCTCTAATG
3475 Left GTTCTCTCTCCAGAGTGCTCTAATG 2480 Right ATGTTCTCTATGGCAAACCTATCAA
3476 Right CATTTTCCTACGATGTTCTCTATGG
KIT Exon12 130-150 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
3481 Left CCTTGTTGTCTTCCTTCCTACAG 3501 Left TGTTGTCTTCCTTCCTACA
3482 Right GCAGTACCATACAGGAACTTAC 3502 Right CATGCAGTACCATACAGGAACTTAC
3483 Left TGTTGTCTTCCTTCCTACAG 3503 Left TTGTTGTCTTCCTTCCTACAG
3484 Right CATGCAGTACCATACAGGAACTTAC 3504 Right ATGCAGTACCATACAGGAACTTA
3485 Left CTTGTTGTCTTCCTTCCTACAG 3505 Left CCTTGTTGTCTTCCTTCCTACAG
3486 Right TGCAGTACCATACAGGAACTTAC 3506 Right GCAGTACCATACAGGAACTT
3487 Left TTGTTGTCTTCCTTCCTACAG 3507 Left TGTTGTCTTCCTTCCTACAG
3488 Right ATGCAGTACCATACAGGAACTTAC 3508 Right CATGCAGTACCATACAGGAACTT
3489 Left CCTTGTTGTCTTCCTTCCTACA 3509 Left ACCTTGTTGTCTTCCTTCCTACAG
3490 Right GCAGTACCATACAGGAACTTAC 3510 Right CAGTACCATACAGGAACTTAC
3491 Left CCTTGTTGTCTTCCTTCCTACAG 3511 Left TTGTTGTCTTCCTTCCTACA
3492 Right GCAGTACCATACAGGAACTTA 3512 Right ATGCAGTACCATACAGGAACTTAC

3493 Left TGTTGTCTTCCTTCCTACAG 3513 Left CTTGTTGTCTTCCTTCCTACAG
3494 Right CATGCAGTACCATACAGGAACTTA 3514 Right TGCAGTACCATACAGGAACTT
3495 Left CTTGTTGTCTTCCTTCCTACAG 3515 Left CCTTGTTGTCTTCCTTCCTACA
3496 Right TGCAGTACCATACAGGAACTTA 3516 Right GCAGTACCATACAGGAACTTA
3497 Left CTTGTTGTCTTCCTTCCTACA 3517 Left TGTTGTCTTCCTTCCTACAG
3498 Right TGCAGTACCATACAGGAACTTAC 3518 Right ATGCAGTACCATACAGGAACTTAC
3499 Left TTGTTGTCTTCCTTCCTACAG 3519 Left TTGTTGTCTTCCTTCCTACAG
3500 Right TGCAGTACCATACAGGAACTTAC 3520 Right ATGCAGTACCATACAGGAACTT
KIT Exon12 151-200 bases Seq. . 2527 Left TTACCTTGTTGTCTTCCTTCCTACA

Primer Sequence ID 2528 Right GCATGCAGTACCATACAGGAACTTA
3521 Left TACCTTGTTGTCTTCCTTCCTACAG ____ 3522 Right CATGCAGTACCATACAGGAACTTAC 2529 Left ACCTTGTTGTCTTCCTTCCTACAG
Seq. . 2530 Right CATGCAGTACCATACAGGAACTTAC
ID Primer Sequence 3523 Left TTACCTTGTTGTCTTCCTTCCTACA 2531 Left ACCACTTACCTTGTTGTCTTCCTTC
3524 Right CATGCAGTACCATACAGGAACTTAC 2532 Right CATGCAGTACCATACAGGAACTTAC
3525 Left TACCTTGTTGTCTTCCTTCCTACAG _____ 3526 Right GCATGCAGTACCATACAGGAACTTA 2533 Left CTTACCTTGTTGTCTTCCTTCCTAC
2534 Right CATGCAGTACCATACAGGAACTTAC

2548 Right CATGCAGTACCATACAGGAACTTAC
3535 Left ACTTACCTTGTTGTCTTCCTTCCTA
3536 Right CATGCAGTACCATACAGGAACTTAC 2549 Left CATCACCACTTACCTTGTTGTCTT
2550 Right CATGCAGTACCATACAGGAACTTAC
3537 Left CATCACCACTTACCTTGTTGTCTTC
3538 Right CATGCAGTACCATACAGGAACTTAC 2551 Left ACCTTGTTGTCTTCCTTCCTACAG
2552 Right GCATGCAGTACCATACAGGAACTTA
3539 Left CACTTACCTTGTTGTCTTCCTTCCT
3540 Right CATGCAGTACCATACAGGAACTTAC 2553 Left ACCACTTACCTTGTTGTCTTCCTTC
2554 Right GCATGCAGTACCATACAGGAACTTA
3541 Left CCACTTACCTTGTTGTCTTCCTTC
3542 Right CATGCAGTACCATACAGGAACTTAC 2555 Left TACCTTGTTGTCTTCCTTCCTACA
2556 Right CATGCAGTACCATACAGGAACTTAC
3543 Left CACTTACCTTGTTGTCTTCCTTCC
3544 Right CATGCAGTACCATACAGGAACTTAC 2557 Left CTTACCTTGTTGTCTTCCTTCCTAC
2558 Right GCATGCAGTACCATACAGGAACTTA
3545 Left ACTTACCTTGTTGTCTTCCTTCCTAC
3546 Right CATGCAGTACCATACAGGAACTTAC 2559 Left ACTTACCTTGTTGTCTTCCTTCCTA
2560 Right GCATGCAGTACCATACAGGAACTTA
3547 Left ACCACTTACCTTGTTGTCTTCCTT
KIT Exon12 201-300 bases S eq. .
Primer S equence ID 581 Left GCCATAGAGAACATCGTAGGAAAAT
3561 Left CTTTTGATAGGTTTGCCATAGAGAA 2582 Right CATGCAGTACCATACAGGAACTTAC
3562 Right CATGCAGTACCATACAGGAACTTAC ______ 2583 Left TTGATAGGTTTGCCATAGAGAACA
3563 Left AGAACATCGTAGGAAAATGTCTCTG 2584 Right CATGCAGTACCATACAGGAACTTAC
3564 Right CATGCAGTACCATACAGGAACTTAC
2585 Left AATTCCTTTATTGATTTTGAAACTGC
3565 Left CCATAGAGAACATCGTAGGAAAATG 2586 Right CATGCAGTACCATACAGGAACTTAC
3566 Right CATGCAGTACCATACAGGAACTTAC
2587 Left TTTGATAGGTTTGCCATAGAGAACA
3567 Left CTTTTGATAGGTTTGCCATAGAGAA 2588 Right CATGCAGTACCATACAGGAACTTAC
3568 Right GCATGCAGTACCATACAGGAACTTA
2589 Left TCTCTGGACAACATTGTTTTTAATTC
3569 Left TCCTTTATTGATTTTGAAACTGCAC 2590 Right CATGCAGTACCATACAGGAACTTAC
3570 Right CATGCAGTACCATACAGGAACTTAC
2591 Left TAGGTTTGCCATAGAGAACATCGTA
3571 Left TGATAGGTTTGCCATAGAGAACATC 2592 Right CATGCAGTACCATACAGGAACTTAC
3572 Right CATGCAGTACCATACAGGAACTTAC
__________________________________________ 2593 Left TGTCTCTGGACAACATTGTTTTTAAT
3573 Left AGAACATCGTAGGAAAATGTCTCTG 2594 Right CATGCAGTACCATACAGGAACTTAC
3574 Right GCATGCAGTACCATACAGGAACTTA
2595 Left AATGTCTCTGGACAACATTGTTTTTA
3575 Left CCATAGAGAACATCGTAGGAAAATG 2596 Right CATGCAGTACCATACAGGAACTTAC
3576 Right GCATGCAGTACCATACAGGAACTTA _____ S eq. . 2597 Left TGGACAACATTGTTTTTAATTCCTT
Pnmer S equence ID 598 Right GCATTTTAGCAAAAAGCACAACT

3577 Left ATTCCTTTATTGATTTTGAAACTGC _______ 3578 Right CATGCAGTACCATACAGGAACTTAC 2599 Left TCCTTTATTGATTTTGAAACTGCAC
2600 Right GCATGCAGTACCATACAGGAACTTA
3579 Left ATGTCTCTGGACAACATTGTTTTTA
3580 Right CATGCAGTACCATACAGGAACTTAC

KIT Exon12 301-400 bases Seq. S eq.
Primer Sequence Primer Sequence ID ID
3601 Left TGAAACAATGAGTTTTCTGTGAAAT 2621 Left CCTTTTTGGGTACACATAACAGTGA
3602 Right CATGCAGTACCATACAGGAACTTAC 2622 Right CATGCAGTACCATACAGGAACTTAC
3603 Left CTGAAACAATGAGTTTTCTGTGAAAT 2623 Left CTTTTGATAGGTTTGCCATAGAGAA
3604 Right CATGCAGTACCATACAGGAACTTAC 2624 Right ATTTTAGCAAAAAGCACAACTGG
3605 Left CTTTTTGGGTACACATAACAGTGAC 2625 Left ACCTGAAACAATGAGTTTTCTGTG
3606 Right CATGCAGTACCATACAGGAACTTAC 2626 Right CATGCAGTACCATACAGGAACTTAC
3607 Left TTTTGGGTACACATAACAGTGACTT 2627 Left AGAACATCGTAGGAAAATGTCTCTG
3608 Right CATGCAGTACCATACAGGAACTTAC 2628 Right ATTTTAGCAAAAAGCACAACTGG
3609 Left TTTTTGGGTACACATAACAGTGACTT 2629 Left CCATAGAGAACATCGTAGGAAAATG
3610 Right CATGCAGTACCATACAGGAACTTAC 2630 Right GCATTTTAGCAAAAAGCACAACT
3611 Left CTTTTGATAGGTTTGCCATAGAGAA 2631 Left TGAAACAATGAGTTTTCTGTGAAAT
3612 Right GCATTTTAGCAAAAAGCACAACT 2632 Right GCATGCAGTACCATACAGGAACTTA
3613 Left ACCTGAAACAATGAGTTTTCTGTGA 2633 Left CCTGAAACAATGAGTTTTCTGTGA
3614 Right CATGCAGTACCATACAGGAACTTAC 2634 Right CATGCAGTACCATACAGGAACTTAC
3615 Left AGAACATCGTAGGAAAATGTCTCTG 2635 Left CCATAGAGAACATCGTAGGAAAATG
3616 Right GCATTTTAGCAAAAAGCACAACT 2636 Right ATTTTAGCAAAAAGCACAACTGG
3617 Left AAACAATGAGTTTTCTGTGAAATTG 2637 Left GTTCCACCTGAAACAATGAGTTTT
3618 Right CATGCAGTACCATACAGGAACTTAC 2638 Right CATGCAGTACCATACAGGAACTTAC
3619 Left GAAACAATGAGTTTTCTGTGAAATTG 2639 Left CTGAAACAATGAGTTTTCTGTGAAAT
3620 Right CATGCAGTACCATACAGGAACTTAC 2640 Right GCATGCAGTACCATACAGGAACTTA
KIT Exon13 131-150 bases Seq. . 2649 Left TGCATGTTTCCAATTTTAG
Primer Sequence ID 650 Right CAGCTTGGACACGGCTTT
3641 Left TGCATGTTTCCAATTTTAG
3642 Right CAGCTTGGACACGGCTTTAC 2651 Left TGCATGTTTCCAATTTTAG
__________________________________________ 2652 Right AGCTTGGACACGGCTTTAC
3643 Left AATGCATGTTTCCAATTTTAG
3644 Right GCTTGGACACGGCTTTAC
3645 Left TGCATGTTTCCAATTTTAG 2653 Left AATGCATGTTTCCAATTTTA
3646 Right CAGCTTGGACACGGCTTTA 2654 Right GCTTGGACACGGCTTTAC
3655 Left ATGCATGTTTCCAATTTTAG
3656 Right AGCTTGGACACGGCTTTA
Seq. .
Primer Sequence ID 3657 Left TGCATGTTTCCAATTTTA
3647 Left ATGCATGTTTCCAATTTTAG 3658 Right CAGCTTGGACACGGCTTTAC
3648 Right AGCTTGGACACGGCTTTAC

3659 Left ATGCATGTTTCCAATTTTAG 3670 Right AGCTTGGACACGGCTTTA
3660 Right GCTTGGACACGGCTTTAC
3671 Left ATGCATGTTTCCAATTTT
3661 Left TGCATGTTTCCAATTTTA 3672 Right AGCTTGGACACGGCTTTAC
3662 Right CAGCTTGGACACGGCTTTA
3673 Left GCATGTTTCCAATTTTAG
3663 Left ATGCATGTTTCCAATTTTA 3674 Right CAGCTTGGACACGGCTTTAC
3664 Right AGCTTGGACACGGCTTTAC
3675 Left TGCATGTTTCCAATTTTAG
3665 Left AATGCATGTTTCCAATTTT 3676 Right GCTTGGACACGGCTTTAC
3666 Right GCTTGGACACGGCTTTAC
3677 Left GCATGTTTCCAATTTTAG
3667 Left TGCATGTTTCCAATTTTA 3678 Right CAGCTTGGACACGGCTTTA
3668 Right CAGCTTGGACACGGCTTT
3679 Left TGCATGTTTCCAATTTTA
3669 Left TGCATGTTTCCAATTTTAG 3680 Right AGCTTGGACACGGCTTTAC
KIT Exon13 151-200 bases Seq. .
Pnmer Sequence ID 3701 Left TGCTAAAATGCATGTTTCCAAT
3681 Left TGCTAAAATGCATGTTTCCAAT 3702 Right TGTTTTGATAACCTGACAGACAATAA
3682 Right CATGTTTTGATAACCTGACAGACAA
3703 Left TAAAATGCATGTTTCCAATTTTAG
3683 Left TAAAATGCATGTTTCCAATTTTAG 3704 Right ATGTTTTGATAACCTGACAGACAAT
3684 Right CATGTTTTGATAACCTGACAGACAA
3705 Left TGCTAAAATGCATGTTTCCAAT
3685 Left TGCTAAAATGCATGTTTCCAAT 3706 Right CATGTTTTGATAACCTGACAGACA
3686 Right TTGATAACCTGACAGACAATAAAAGG
3707 Left TAAAATGCATGTTTCCAATTTTAG
3687 Left TAAAATGCATGTTTCCAATTTTAG 3708 Right TGTTTTGATAACCTGACAGACAATAA
3688 Right TTGATAACCTGACAGACAATAAAAGG
__________________________________________ 3709 Left TAAAATGCATGTTTCCAATTTTAG

3689 Left TGCTAAAATGCATGTTTCCAAT 3710 Right CATGTTTTGATAACCTGACAGACA
3690 Right TGATAACCTGACAGACAATAAAAGG ______ 3711 Left TGCTAAAATGCATGTTTCCAAT
3691 Left TAAAATGCATGTTTCCAATTTTAG 3712 Right CTGACAGACAATAAAAGGCAGCTT
3692 Right TGATAACCTGACAGACAATAAAAGG
3713 Left TAAAATGCATGTTTCCAATTTTAG
3693 Left AAAATGCATGTTTCCAATTTTAG 3714 Right CTGACAGACAATAAAAGGCAGCTT
3694 Right CATGTTTTGATAACCTGACAGACAA
3715 Left TGCTAAAATGCATGTTTCCAAT
3695 Left TGCTAAAATGCATGTTTCCAAT 3716 Right TGTTTTGATAACCTGACAGACAATA
3696 Right CATGTTTTGATAACCTGACAGACAAT
3697 Left TAAAATGCATGTTTCCAATTTTAG
3698 Right CATGTTTTGATAACCTGACAGACAAT 3717 Left TAAAATGCATGTTTCCAATTTTAG
3718 Right TGTTTTGATAACCTGACAGACAATA
Seq. . 3719 Left AAAATGCATGTTTCCAATTTTAG
Pnmer Sequence ID 3720 Right TTGATAACCTGACAGACAATAAAAGG
3699 Left TGCTAAAATGCATGTTTCCAAT
3700 Right ATGTTTTGATAACCTGACAGACAAT

KIT Exon13 201-300 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
3721 Left CTTGACATCAGTTTGCCAGTTGT 3 741 Left TAAAATGCATGTTTCCAATTTTAG

3722 Right GAGAGAACAACAGTCTGGGTAAAAA 3 742 Right ACAATGAGGAAAACAAAATCTAGCA
3723 Left TGCTAAAATGCATGTTTCCAAT 3 743 Left CTTGACATCAGTTTGCCAGTTG
3724 Right GAGAGAACAACAGTCTGGGTAAAAA 3 744 Right AGAGAGAACAACAGTCTGGGTAAAA
3725 Left TAAAATGCATGTTTCCAATTTTAG 2745 Left CTTGACATCAGTTTGCCAGTTG
3726 Right GAGAGAACAACAGTCTGGGTAAAAA 3 746 Right AAGAGAGAACAACAGTCTGGGTAAA
3727 Left TTGACATCAGTTTGCCAGTTGT 3 747 Left TGCTAAAATGCATGTTTCCAAT
3728 Right GAGAGAACAACAGTCTGGGTAAAAA 3 748 Right GGAAAACAAAATCTAGCAAGAGAGA
3729 Left CTTGACATCAGTTTGCCAGTTGT 3 749 Left TGCTAAAATGCATGTTTCCAAT
3730 Right AGAGAGAACAACAGTCTGGGTAAAA 2750 Right GAGGAAAACAAAATCTAGCAAGAGA
3731 Left TGCTAAAATGCATGTTTCCAAT 3 751 Left CTTGACATCAGTTTGCCAGTTG
3732 Right AGAGAGAACAACAGTCTGGGTAAAA 3 752 Right CATGTTTTGATAACCTGACAGACAA
3733 Left TGCTAAAATGCATGTTTCCAAT 3 753 Left TAAAATGCATGTTTCCAATTTTAG

3734 Right AAGAGAGAACAACAGTCTGGGTAAA 3 754 Right GGAAAACAAAATCTAGCAAGAGAGA
3735 Left TAAAATGCATGTTTCCAATTTTAG 2755 Left TAAAATGCATGTTTCCAATTTTAG
3736 Right AAGAGAGAACAACAGTCTGGGTAAA 3 756 Right GAGGAAAACAAAATCTAGCAAGAGA
3737 Left CTTGACATCAGTTTGCCAGTTG 3 757 Left ATCAGTTTGCCAGTTGTGCTTT
3738 Right GAGAGAACAACAGTCTGGGTAAAAA 3 758 Right GAGAGAACAACAGTCTGGGTAAAAA
3739 Left TTGACATCAGTTTGCCAGTTGT 3 759 Left ATCAGTTTGCCAGTTGTGCTT
3740 Right AGAGAGAACAACAGTCTGGGTAAAA 3 760 Right GAGAGAACAACAGTCTGGGTAAAAA
KIT Exon13 301-400 bases Seq. 3 771 Left CTTGACATCAGTTTGCCAGTTGT
Primer Sequence ID 3 772 Right GCTGTTCTACCCCATAATGATAAAA
3761 Left CTTGACATCAGTTTGCCAGTTGT
3762 Right ACGACAATAACTAGGGTATGTCCTG 3 773 Left TGCTAAAATGCATGTTTCCAAT
3 774 Right GCTGATGTCGTTGATGTTACAAATA
3763 Left TGCTAAAATGCATGTTTCCAAT
3764 Right ACGACAATAACTAGGGTATGTCCTG 3 775 Left TGCTAAAATGCATGTTTCCAAT
3 776 Right GCTGTTCTACCCCATAATGATAAAA
3765 Left TGCTAAAATGCATGTTTCCAAT
3766 Right TCGTTGATGTTACAAATACGACAAT 3 777 Left TAAAATGCATGTTTCCAATTTTAG
3 778 Right GCTGATGTCGTTGATGTTACAAATA
3767 Left TAAAATGCATGTTTCCAATTTTAG _____ 3768 Right ACGACAATAACTAGGGTATGTCCTG ¨3 779 Left TTGACATCAGTTTGCCAGTTGT
3 780 Right ACGACAATAACTAGGGTATGTCCTG
3769 Left TAAAATGCATGTTTCCAATTTTAG
3770 Right TCGTTGATGTTACAAATACGACAAT 3 781 Left TTGACATCAGTTTGCCAGTTGT
Seq. 3 782 Right GCTGTTCTACCCCATAATGATAAAA
ID Primer Sequence 3783 Left CTTGACATCAGTTTGCCAGTTG 3 792 Right ACAATGAGGAAAACAAAATCTAGCA
3784 Right ACGACAATAACTAGGGTATGTCCTG
3 793 Left CTTGACATCAGTTTGCCAGTTG
3785 Left CTTGACATCAGTTTGCCAGTTGT 3 794 Right GAGGAAAACAAAATCTAGCAAGAGA
3786 Right GAGGAAAACAAAATCTAGCAAGAGA
3 795 Left CTTGACATCAGTTTGCCAGTTG
3787 Left CTTGACATCAGTTTGCCAGTTG 3 796 Right GGAAAACAAAATCTAGCAAGAGAGA
3788 Right GCTGTTCTACCCCATAATGATAAAA
3 797 Left ATCAGTTTGCCAGTTGTGCTTT
3789 Left TTGACATCAGTTTGCCAGTTGT 3 798 Right ACGACAATAACTAGGGTATGTCCTG
3790 Right GAGGAAAACAAAATCTAGCAAGAGA
3 799 Left ATCAGTTTGCCAGTTGTGCTTT
3791 Left CTTGACATCAGTTTGCCAGTTG 3 800 Right GCTGTTCTACCCCATAATGATAAAA
KIT Exon10-11 301-400 bases Seq.
Primer Sequence Primer Sequence ID ID
3801 Left GATTCCACATTTCTCTTCCATTGTA 3 821 Left AGTTTGTGATTCCACATTTCTCTTC
3802 Right TTATGTGTACCCAAAAAGGTGACAT 3 822 Right GAAAGCCCCTGTTTCATACTGAC
3803 Left GATTCCACATTTCTCTTCCATTGTA 3 823 Left ATTCCACATTTCTCTTCCATTGTA
3804 Right TATGTGTACCCAAAAAGGTGACAT 3 824 Right GTTATGTGTACCCAAAAAGGTGACA
3805 Left GATTCCACATTTCTCTTCCATTGTA 2825 Left ATTCCACATTTCTCTTCCATTGTA
3806 Right TTATGTGTACCCAAAAAGGTGACA 3 826 Right TATGTGTACCCAAAAAGGTGACAT
3807 Left GATTCCACATTTCTCTTCCATTGT 3 827 Left AGTTTGTGATTCCACATTTCTCTTC
3808 Right TTATGTGTACCCAAAAAGGTGACAT 3 828 Right AAAGCCCCTGTTTCATACTGAC
3809 Left ATTCCACATTTCTCTTCCATTGTA 3 829 Left TGATTCCACATTTCTCTTCCATT
3810 Right TTATGTGTACCCAAAAAGGTGACAT 3 830 Right TATGTGTACCCAAAAAGGTGACAT
3811 Left GATTCCACATTTCTCTTCCATTG 3 831 Left GATTCCACATTTCTCTTCCATTG
3812 Right TTATGTGTACCCAAAAAGGTGACAT 3 832 Right TATGTGTACCCAAAAAGGTGACAT
3813 Left GATTCCACATTTCTCTTCCATTGT 3 833 Left ATTCCACATTTCTCTTCCATTGTA
3814 Right TATGTGTACCCAAAAAGGTGACAT 3 834 Right TTATGTGTACCCAAAAAGGTGACA
3815 Left GATTCCACATTTCTCTTCCATTGTA 2835 Left GATTCCACATTTCTCTTCCATTG
3816 Right ATGTGTACCCAAAAAGGTGACAT 3 836 Right TTATGTGTACCCAAAAAGGTGACA
3817 Left GATTCCACATTTCTCTTCCATTGT 3 837 Left AAGTTTGTGATTCCACATTTCTCTT
3818 Right TTATGTGTACCCAAAAAGGTGACA 3 838 Right GAAAGCCCCTGTTTCATACTGAC
3819 Left GATTCCACATTTCTCTTCCATTGTA 3 839 Left AAAGTTTGTGATTCCACATTTCTCT
3820 Right TATGTGTACCCAAAAAGGTGACA 3 840 Right GAAAGCCCCTGTTTCATACTGAC
KIT Exon10-11 401-500 bases S ea ID
ID' Primer Sequence 2843 Left AGTTTGTGATTCCACATTTCTCTTC
3841 Left AGTTTGTGATTCCACATTTCTCTTC 2844 Right GCAATTTCACAGAAAACTCATTGTT
3842 Right TTATGTGTACCCAAAAAGGTGACAT 2845 Left AGTTTGTGATTCCACATTTCTCTTC
S eq. Primer Sequence 2846 Right ATTTCACAGAAAACTCATTGTTTCA

2 864 Right CTGTTATGTGTACCCAAAAAGGTG
3847 Left AAGTTTGTGATTCCACATTTCTCTT
3848 Right TTATGTGTACCCAAAAAGGTGACAT 2865 Left AGTTTGTGATTCCACATTTCTCTTC
2866 Right TCACAGAAAACTCATTGTTTCAGGT
3849 Left AAAGTTTGTGATTCCACATTTCTCT
3850 Right TTATGTGTACCCAAAAAGGTGACAT 2867 Left AGTTTGTGATTCCACATTTCTCTTC
2868 Right AATTTCACAGAAAACTCATTGTTTCA
3851 Left AAGTTTGTGATTCCACATTTCTCTT
3852 Right GCAATTTCACAGAAAACTCATTGTT 2869 Left AAGTTTGTGATTCCACATTTCTCTT
2870 Right ATTTCACAGAAAACTCATTGTTTCA
3853 Left AAAGTTTGTGATTCCACATTTCTCT
3854 Right GCAATTTCACAGAAAACTCATTGTT 2871 Left AAAGTTTGTGATTCCACATTTCTCT
2872 Right ATTTCACAGAAAACTCATTGTTTCA
3855 Left AGTTTGTGATTCCACATTTCTCTTC
3856 Right GTTATGTGTACCCAAAAAGGTGACA 2873 Left AGTTTGTGATTCCACATTTCTCTTC
2 874 Right CTGTTATGTGTACCCAAAAAGGTGA
3857 Left AGTTTGTGATTCCACATTTCTCTTC
3858 Right TGTTATGTGTACCCAAAAAGGTGAC 2875 Left GATTCCACATTTCTCTTCCATTGTA
2876 Right ATTTCACAGAAAACTCATTGTTTCA
3859 Left GATTCCACATTTCTCTTCCATTGTA
3860 Right TTATGTGTACCCAAAAAGGTGACAT 2877 Left AGTTTGTGATTCCACATTTCTCTTC
2878 Right ACAGAAAACTCATTGTTTCAGGTG
3861 Left GATTCCACATTTCTCTTCCATTGTA
3862 Right GCAATTTCACAGAAAACTCATTGTT 2879 Left AGTTTGTGATTCCACATTTCTCTTC
2880 Right CACAGAAAACTCATTGTTTCAGGT
3863 Left AGTTTGTGATTCCACATTTCTCTTC
KIT Exon10-11 501-600 bases S eq. Primer ID
S equence ID 3 897 Left AGTTTGTGATTCCACATTTCTCTTC
3881 Left AGTTTGTGATTCCACATTTCTCTTC 3 898 Right TTGTGCAGTTTCAAAATCAATAAAG
3882 Right TTCTCTATGGCAAACCTATCAAAAG
3 899 Left AGTTTGTGATTCCACATTTCTCTTC
3883 Left AGTTTGTGATTCCACATTTCTCTTC 3 900 Right GGAATTAAAAACAATGTTGTCCAGA
3884 Right GTTCTCTATGGCAAACCTATCAAAA
3 901 Left AAGTTTGTGATTCCACATTTCTCTT
3885 Left AGTTTGTGATTCCACATTTCTCTTC 3 902 Right TTCTCTATGGCAAACCTATCAAAAG
3886 Right ATGTTGTCCAGAGACATTTTCCTAC
3 903 Left AAAGTTTGTGATTCCACATTTCTCT
3887 Left AGTTTGTGATTCCACATTTCTCTTC 3 904 Right TTCTCTATGGCAAACCTATCAAAAG
3888 Right CATTTTCCTACGATGTTCTCTATGG
__________________________________________ 3 905 Left AAGTTTGTGATTCCACATTTCTCTT
3889 Left AGTTTGTGATTCCACATTTCTCTTC 3 906 Right GTTCTCTATGGCAAACCTATCAAAA
3890 Right ATGTTCTCTATGGCAAACCTATCAA
3 907 Left AAAGTTTGTGATTCCACATTTCTCT
3891 Left AGTTTGTGATTCCACATTTCTCTTC 3 908 Right GTTCTCTATGGCAAACCTATCAAAA
3892 Right CATTTGTGCAGTTTCAAAATCAATA ____ 3 909 Left GATTCCACATTTCTCTTCCATTGTA
3893 Left AGTTTGTGATTCCACATTTCTCTTC 3 910 Right TTCTCTATGGCAAACCTATCAAAAG
3894 Right AATGTTGTCCAGAGACATTTTCCTA
3 911 Left GATTCCACATTTCTCTTCCATTGTA
3895 Left AGTTTGTGATTCCACATTTCTCTTC 3 912 Right GTTCTCTATGGCAAACCTATCAAAA
3896 Right AGGAATTAAAAACAATGTTGTCCAG
S eq. Primer Sequence 3913 Left AAGTTTGTGATTCCACATTTCTCTT 3 917 Left GATTCCACATTTCTCTTCCATTGTA
3914 Right ATGTTGTCCAGAGACATTTTCCTAC 3 918 Right ATGTTGTCCAGAGACATTTTCCTAC
3915 Left AAAGTTTGTGATTCCACATTTCTCT 3 919 Left AAGTTTGTGATTCCACATTTCTCTT
3916 Right ATGTTGTCCAGAGACATTTTCCTAC 3 920 Right CATTTTCCTACGATGTTCTCTATGG
KIT Exon10-11 601-800 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
3921 Left TCTATTCTGCAGTATTGTGGTTTCA 3 941 Left TCTATTCTGCAGTATTGTGGTTTCA
3922 Right TTCTCTATGGCAAACCTATCAAAAG 3 942 Right AATGTTGTCCAGAGACATTTTCCTA
3923 Left TCTGCAGTATTGTGGTTTCAAGTTA 3 943 Left TCTGCAGTATTGTGGTTTCAAGTTA
3924 Right TTCTCTATGGCAAACCTATCAAAAG 3 944 Right CATTTTCCTACGATGTTCTCTATGG
3925 Left TCTATTCTGCAGTATTGTGGTTTCA 2945 Left TCTGCAGTATTGTGGTTTCAAGTTA
3926 Right GTTCTCTATGGCAAACCTATCAAAA 3 946 Right ATGTTCTCTATGGCAAACCTATCAA
3927 Left TCTGCAGTATTGTGGTTTCAAGTTA 3 947 Left GTACAATGTAACCAAGGTGAAGCTC
3928 Right GTTCTCTATGGCAAACCTATCAAAA 3 948 Right TTCTCTATGGCAAACCTATCAAAAG
3929 Left TCTATTCTGCAGTATTGTGGTTTCA 3 949 Left GAGTACAATGTAACCAAGGTGAAGC
3930 Right ATGTTGTCCAGAGACATTTTCCTAC 2950 Right TTCTCTATGGCAAACCTATCAAAAG
3931 Left GGCAGGAATTTGATTGAAGTATAAA 3 951 Left ATTCTGCAGTATTGTGGTTTCAAGT
3932 Right TTCTCTATGGCAAACCTATCAAAAG 3 952 Right TTCTCTATGGCAAACCTATCAAAAG
3933 Left TCTGCAGTATTGTGGTTTCAAGTTA 3 953 Left TCTGCAGTATTGTGGTTTCAAGTTA
3934 Right ATGTTGTCCAGAGACATTTTCCTAC 3 954 Right CATTTGTGCAGTTTCAAAATCAATA
3935 Left GGCAGGAATTTGATTGAAGTATAAA 2955 Left TCTGCAGTATTGTGGTTTCAAGTTA
3936 Right GTTCTCTATGGCAAACCTATCAAAA 3 956 Right AATGTTGTCCAGAGACATTTTCCTA
3937 Left TCTATTCTGCAGTATTGTGGTTTCA 3 957 Left TCTGCAGTATTGTGGTTTCAAGTTA
3938 Right CATTTTCCTACGATGTTCTCTATGG 3 958 Right AGGAATTAAAAACAATGTTGTCCAG
3939 Left TCTATTCTGCAGTATTGTGGTTTCA 3 959 Left GTACAATGTAACCAAGGTGAAGCTC
3940 Right ATGTTCTCTATGGCAAACCTATCAA 3 960 Right GTTCTCTATGGCAAACCTATCAAAA
KIT Exon12-13 301-400 bases Primer Sequence Seq.
Primer Sequence ID ID
3961 Left TACCTTGTTGTCTTCCTTCCTACAG 3967 Left ACCTTGTTGTCTTCCTTCCTACAG
3962 Right CATGTTTTGATAACCTGACAGACAA 3968 Right CATGTTTTGATAACCTGACAGACAA
3963 Left TTACCTTGTTGTCTTCCTTCCTACA 3969 Left TACCTTGTTGTCTTCCTTCCTACAG
3964 Right CATGTTTTGATAACCTGACAGACAA 3970 Right TGATAACCTGACAGACAATAAAAGG
3965 Left TACCTTGTTGTCTTCCTTCCTACAG 3971 Left ACCACTTACCTTGTTGTCTTCCTTC
3966 Right TTGATAACCTGACAGACAATAAAAGG3972 Right CATGTTTTGATAACCTGACAGACAA

3973 Left CTTACCTTGTTGTCTTCCTTCCTAC 3987 Left CCACTTACCTTGTTGTCTTCCTTC
3974 Right CATGTTTTGATAACCTGACAGACAA 3988 Right CATGTTTTGATAACCTGACAGACAA
3975 Left ACTTACCTTGTTGTCTTCCTTCCTA 3989 Left ACTTACCTTGTTGTCTTCCTTCCTAC
3976 Right CATGTTTTGATAACCTGACAGACAA 3990 Right CATGTTTTGATAACCTGACAGACAA
3977 Left CACTTACCTTGTTGTCTTCCTTCCT 3991 Left AC CACTTACCTTGTTGTCTTCCTT
3978 Right CATGTTTTGATAACCTGACAGACAA 3992 Right CATGTTTTGATAACCTGACAGACAA
3979 Left TTACCTTGTTGTCTTCCTTCCTACA 3993 Left TTACCTTGTTGTCTTCCTTCCTACA
3980 Right TTGATAACCTGACAGACAATAAAAGG 3994 Right CATGTTTTGATAACCTGACAGACAAT
3981 Left TACCTTGTTGTCTTCCTTCCTACAG 3995 Left TACCTTGTTGTCTTCCTTCCTACAG
3982 Right CATGTTTTGATAACCTGACAGACAAT 3996 Right ATGTTTTGATAACCTGACAGACAAT
3983 Left TTACCTTGTTGTCTTCCTTCCTACA 3997 Left TACCTTGTTGTCTTCCTTCCTACAG
3984 Right TGATAACCTGACAGACAATAAAAGG 3998 Right TGTTTTGATAACCTGACAGACAATAA
3985 Left CACTTACCTTGTTGTCTTCCTTCC 3999 Left TACCTTGTTGTCTTCCTTCCTACAG
3986 Right CATGTTTTGATAACCTGACAGACAA 4000 Right CATGTTTTGATAACCTGACAGACA
KIT Exon12-13 401-500 bases S eq. . z 020 Right AGAGAGAACAACAGTCTGGGTAAAA
Pnmer Sequence ID Seq.
Primer Sequence 4001 Left TACCTTGTTGTCTTCCTTCCTACAG ID
4002 Right GAGAGAACAACAGTCTGGGTAAAAA z 021 Left TTACCTTGTTGTCTTCCTTCCTACA
z 022 Right AAGAGAGAACAACAGTCTGGGTAAA
4003 Left TACCTTGTTGTCTTCCTTCCTACAG
4004 Right AGAGAGAACAACAGTCTGGGTAAAA z 023 Left TCTGGACAACATTGTTTTTAATTCC
z 024 Right CATGTTTTGATAACCTGACAGACAA
4005 Left TACCTTGTTGTCTTCCTTCCTACAG
4006 Right AAGAGAGAACAACAGTCTGGGTAAA z 025 Left TTACCTTGTTGTCTTCCTTCCTACA
z 026 Right ACAATGAGGAAAACAAAATCTAGCA
4007 Left TGGACAACATTGTTTTTAATTCCTT
4008 Right CATGTTTTGATAACCTGACAGACAA z 027 Left TGGACAACATTGTTTTTAATTCCTT
z 028 Right TTATAATCTAGCATTGCCAAAATCA
4009 Left AGAACATCGTAGGAAAATGTCTCTG
4010 Right CATGTTTTGATAACCTGACAGACAA z 029 Left TACCTTGTTGTCTTCCTTCCTACAG
z 030 Right CAATGAGGAAAACAAAATCTAGCAA
4011 Left TACCTTGTTGTCTTCCTTCCTACAG
4012 Right ACAATGAGGAAAACAAAATCTAGCA z 031 Left TACCTTGTTGTCTTCCTTCCTACAG
z 032 Right AACAATGAGGAAAACAAAATCTAGC
4013 Left TTACCTTGTTGTCTTCCTTCCTACA
4014 Right GAGAGAACAACAGTCTGGGTAAAAA z 033 Left TACCTTGTTGTCTTCCTTCCTACAG
z 034 Right TTATAATCTAGCATTGCCAAAATCA
4015 Left CCATAGAGAACATCGTAGGAAAATG
4016 Right CATGTTTTGATAACCTGACAGACAA z 035 Left TCCTTTATTGATTTTGAAACTGCAC
z 036 Right CATGTTTTGATAACCTGACAGACAA
4017 Left CTGGACAACATTGTTTTTAATTCCT
4018 Right CATGTTTTGATAACCTGACAGACAA z 037 Left TGATAGGTTTGCCATAGAGAACATC
z 038 Right CATGTTTTGATAACCTGACAGACAA
4019 Left TTACCTTGTTGTCTTCCTTCCTACA

4039 Left TGGACAACATTGTTTTTAATTCCTT z 040 Right CAGTTTATAATCTAGCATTGCCAAAA
KIT Exon12-13 501-600 bases Seq.
Primer Sequence Primer Sequence ID ID
4041 Left TACCTTGTTGTCTTCCTTCCTACAG L061 Left AGAACATCGTAGGAAAATGTCTCTG
4042 Right ATGAGATATTCAAGAGGCTGATGTC L062 Right AAGAGAGAACAACAGTCTGGGTAAA
4043 Left TACCTTGTTGTCTTCCTTCCTACAG L063 Left CCATAGAGAACATCGTAGGAAAATG
4044 Right GATGAGATATTCAAGAGGCTGATGT L064 Right GAGAGAACAACAGTCTGGGTAAAAA
4045 Left CTTTTGATAGGTTTGCCATAGAGAA L 0 65 Left TACCTTGTTGTCTTCCTTCCTACAG
4046 Right GAGAGAACAACAGTCTGGGTAAAAA L 0 6 6 Right ACGACAATAACTAGGGTATGTCCTG
4047 Left TGGACAACATTGTTTTTAATTCCTT L 0 6 7 Left CTTTTGATAGGTTTGCCATAGAGAA
4048 Right GAGAGAACAACAGTCTGGGTAAAAA L068 Right CATGTTTTGATAACCTGACAGACAA
4049 Left AGAACATCGTAGGAAAATGTCTCTG L 0 6 9 Left CTGGACAACATTGTTTTTAATTCCT
4050 Right GAGAGAACAACAGTCTGGGTAAAAA L070 Right GAGAGAACAACAGTCTGGGTAAAAA
4051 Left CTTTTGATAGGTTTGCCATAGAGAA L071 Left TACCTTGTTGTCTTCCTTCCTACAG
4052 Right AGAGAGAACAACAGTCTGGGTAAAA L 0 72 Right TCGTTGATGTTACAAATACGACAAT
4053 Left CTTTTGATAGGTTTGCCATAGAGAA L 0 73 Left TACCTTGTTGTCTTCCTTCCTACAG
4054 Right AAGAGAGAACAACAGTCTGGGTAAA L 0 74 Right GCTGATGTCGTTGATGTTACAAATA
4055 Left TGGACAACATTGTTTTTAATTCCTT L 0 75 Left TACCTTGTTGTCTTCCTTCCTACAG
4056 Right AGAGAGAACAACAGTCTGGGTAAAA L076 Right GCTGTTCTACCCCATAATGATAAAA
4057 Left TGGACAACATTGTTTTTAATTCCTT L 0 77 Left TGGACAACATTGTTTTTAATTCCTT
4058 Right AAGAGAGAACAACAGTCTGGGTAAA L 0 7 8 Right ACAATGAGGAAAACAAAATCTAGCA
4059 Left AGAACATCGTAGGAAAATGTCTCTG L079 Left TTACCTTGTTGTCTTCCTTCCTACA
4060 Right AGAGAGAACAACAGTCTGGGTAAAA L080 Right ATGAGATATTCAAGAGGCTGATGTC
KIT Exon12-13 601-800 bases Primer Sequence ID
4081 Left CTTTTGATAGGTTTGCCATAGAGAA Seq. .
4082 Right GATGAGATATTCAAGAGGCTGATGT ID Pnmer Sequence 4091 Left AGAACATCGTAGGAAAATGTCTCTG
4083 Left CTTTTGATAGGTTTGCCATAGAGAA 4092 Right GATGAGATATTCAAGAGGCTGATGT
4084 Right ATGAGATATTCAAGAGGCTGATGTC
4093 Left AGAACATCGTAGGAAAATGTCTCTG
4085 Left TGGACAACATTGTTTTTAATTCCTT 4094 Right TCAGTGGCTACATATGATCAAGAAA
4086 Right TTTCAGTGGCTACATATGATCAAGA
4095 Left AGAACATCGTAGGAAAATGTCTCTG
4087 Left TGGACAACATTGTTTTTAATTCCTT 4096 Right TTCAGTGGCTACATATGATCAAGAA
4088 Right TTCAGTGGCTACATATGATCAAGAA
4097 Left AGAACATCGTAGGAAAATGTCTCTG
4089 Left AGAACATCGTAGGAAAATGTCTCTG 4098 Right TTTCAGTGGCTACATATGATCAAGA
4090 Right ATGAGATATTCAAGAGGCTGATGTC

4099 Left TACCTTGTTGTCTTCCTTCCTACAG 4109 Left TACCTTGTTGTCTTCCTTCCTACAG
4100 Right TCAGTGGCTACATATGATCAAGAAA Al 10 Right ATGTTTGCCTCATTTGGTGTATATT
__________________________________________ 4111 Left ACAGTGACTTTAAGGAACTCCAGTG
4112 Right GAGAGAACAACAGTCTGGGTAAAAA
4101 Left TACCTTGTTGTCTTCCTTCCTACAG _____ 4102 Right TTCAGTGGCTACATATGATCAAGAA 4113 Left CCATAGAGAACATCGTAGGAAAATG
__________________________________________ 4114 Right GATGAGATATTCAAGAGGCTGATGT
4103 Left TACCTTGTTGTCTTCCTTCCTACAG _____ ¨
4104 Right TTTCAGTGGCTACATATGATCAAGA 4115 Left CCATAGAGAACATCGTAGGAAAATG
__________________________________________ 4116 Right ATGAGATATTCAAGAGGCTGATGTC
4105 Left ACAGTGACTTTAAGGAACTCCAGTG _______ 4106 Right GATGAGATATTCAAGAGGCTGATGT 4117 Left TACCTTGTTGTCTTCCTTCCTACAG
__________________________________________ 4118 Right AAGGCTTCAAATTGGAAACTTATTT
4107 Left ACAGTGACTTTAAGGAACTCCAGTG _______ 4108 Right ATGAGATATTCAAGAGGCTGATGTC 4119 Left CTTTTGATAGGTTTGCCATAGAGAA
4120 Right ACGACAATAACTAGGGTATGTCCTG
KIT Exon10-13 801-1000 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
4121 Left AAGTTTGTGATTCCACATTTCTCTT 4141 Left CAAAGTTTGTGATTCCACATTTCT
4122 Right CATGTTTTGATAACCTGACAGACAA 4142 Right CATGTTTTGATAACCTGACAGACAA
4123 Left GATTCCACATTTCTCTTCCATTGTA 4143 Left GTGATTCCACATTTCTCTTCCATT
4124 Right CATGTTTTGATAACCTGACAGACAA 4144 Right CATGTTTTGATAACCTGACAGACAA
4125 Left AGTTTGTGATTCCACATTTCTCTTC 4145 Left AAAGTTTGTGATTCCACATTTCTCTT
4126 Right TTGATAACCTGACAGACAATAAAAGG4146 Right CATGTTTTGATAACCTGACAGACAA
4127 Left AGTTTGTGATTCCACATTTCTCTTC 4147 Left AGTTTGTGATTCCACATTTCTCTTC
4128 Right TGATAACCTGACAGACAATAAAAGG 4148 Right ATGTTTTGATAACCTGACAGACAAT
4129 Left CAAAGTTTGTGATTCCACATTTCTC 4149 Left AGTTTGTGATTCCACATTTCTCTTC
4130 Right CATGTTTTGATAACCTGACAGACAA 4150 Right TGTTTTGATAACCTGACAGACAATAA
4131 Left AGTTTGTGATTCCACATTTCTCTTC 4151 Left AGTTTGTGATTCCACATTTCTCTTC
4132 Right CATGTTTTGATAACCTGACAGACAAT 4152 Right CATGTTTTGATAACCTGACAGACA
4133 Left AAGTTTGTGATTCCACATTTCTCTT 4153 Left AAGTTTGTGATTCCACATTTCTCTT
4134 Right TTGATAACCTGACAGACAATAAAAGG4154 Right CATGTTTTGATAACCTGACAGACAAT
4135 Left AAAGTTTGTGATTCCACATTTCTCT 4155 Left AGTTTGTGATTCCACATTTCTCTTC
4136 Right TTGATAACCTGACAGACAATAAAAGG4156 Right CTGACAGACAATAAAAGGCAGCTT
4137 Left AAGTTTGTGATTCCACATTTCTCTT 4157 Left AGTTTGTGATTCCACATTTCTCTTC
4138 Right TGATAACCTGACAGACAATAAAAGG 4158 Right TGTTTTGATAACCTGACAGACAATA
4139 Left AAAGTTTGTGATTCCACATTTCTCT 4159 Left CAAAGTTTGTGATTCCACATTTCTC
4140 Right TGATAACCTGACAGACAATAAAAGG 4160 Right TTGATAACCTGACAGACAATAAAAGG
KIT Exon10-13 2kb Seq. Primer Sequence ID

4161 Left AATATAAACCGTCCATAAAGGAAGC z 181 Left AGTTTGTGATTCCACATTTCTCTTC
4162 Right ATTATGGAAATAATGAAGGCACAAA z 182 Right GAATCCTCTGGTTCACTTCTGTTTA
S eq. .
Pnmer S equence ID z 183 Left TCTATTCTGCAGTATTGTGGTTTCA
4163 Left AATATAAACCGTCCATAAAGGAAGC z 184 Right GAATCCTCTGGTTCACTTCTGTTTA
4164 Right TTCTGAGCGCTACTAGTTGAAAAAT
z 185 Left AGAATCTTCCATGTTTTTCAGACAG
4165 Left AATATAAACCGTCCATAAAGGAAGC z 186 Right GACTGGAAAGTGAGTGAAAACCTAA
4166 Right ATTTATGGGGATTATTGGAAGACAT
z 187 Left AATATAAACCGTCCATAAAGGAAGC
4167 Left TCTATTCTGCAGTATTGTGGTTTCA z 188 Right ACAAGACTGGAAAGTGAGTGAAAAC
4168 Right TTCTGAGCGCTACTAGTTGAAAAAT
z 189 Left TTTTAACACTTTGCCAGACACTGTA
4169 Left AATATAAACCGTCCATAAAGGAAGC z 190 Right GATGAGATATTCAAGAGGCTGATGT
4170 Right CTGGAAAGTGAGTGAAAACCTAAAA
z 191 Left TTTTAACACTTTGCCAGACACTGTA
4171 Left AATATAAACCGTCCATAAAGGAAGC z 192 Right ATGAGATATTCAAGAGGCTGATGTC
4172 Right GACTGGAAAGTGAGTGAAAACCTAA
z 193 Left GGATGTTTAGGCTCTGTCTACCATA
4173 Left AGTTTGTGATTCCACATTTCTCTTC z 194 Right GAGAGAACAACAGTCTGGGTAAAAA
4174 Right GATTAATTCTGCCTCCCATAAAAAT
z 195 Left TCTGCAGTATTGTGGTTTCAAGTTA
4175 Left ACTGCCTTTATGAAGTCATGTTAGC z 196 Right GAATCCTCTGGTTCACTTCTGTTTA
4176 Right ATGAGATATTCAAGAGGCTGATGTC
z 197 Left AATATAAACCGTCCATAAAGGAAGC
4177 Left TCTGCAGTATTGTGGTTTCAAGTTA z 198 Right TATGGAAATAATGAAGGCACAAACT
4178 Right TTCTGAGCGCTACTAGTTGAAAAAT
z 199 Left AATACATGCATCACACCATACTGTC
4179 Left AGTTTGTGATTCCACATTTCTCTTC L200 Right GAGAGAACAACAGTCTGGGTAAAAA
4180 Right AATATTCAGCTTCAGTGGATCAGAC
KIT Exon10-13 5kb S eq. . 4213 Left ACTGCCTTTATGAAGTCATGTTAGC

Primer Sequence ID 4214 Right CAGAAACTAGGTGTCCTTTTTGTGT
4201 Left GGATGTTTAGGCTCTGTCTACCATA
4202 Right CAGAAACTAGGTGTCCTTTTTGTGT
_________________________________________ Seq. .
S
4203 Left GGATGTTTAGGCTCTGTCTACCATA ID Primer equence 4204 Right ACCAATAATTTGGATGATTTTCTGA 4215 Left AATACATGCATCACACCATACTGTC
_________________________________________ 4216 Right TAATTTGGATGATTTTCTGAACCAT
4205 Left GGATGTTTAGGCTCTGTCTACCATA _____ 4206 Right TAATTTGGATGATTTTCTGAACCAT 4217 Le ft AATACATGCATCACACCATACTGTC
4218 Right ACCAATAATTTGGATGATTTTCTGA
4207 Left GGATGTTTAGGCTCTGTCTACCATA _____ 4208 Right TCTGATTAGCATAGAAACACCATGA 4219 Le ft AATACATGCATCACACCATACTGTC
4220 Right TCTGATTAGCATAGAAACACCATGA
4209 Left GGATGTTTAGGCTCTGTCTACCATA _____ 4210 Right GGTATCCCGTATTACCTCAAACTCT 7221 Left AATACATGCATCACACCATACTGTC
4222 Right GGTATCCCGTATTACCTCAAACTCT
4211 Left GGATGTTTAGGCTCTGTCTACCATA _____ 4212 Right ACTTCCTGCTTCTGATTAGCATAGA 4223 Left AATACATGCATCACACCATACTGTC
4224 Right ACTTCCTGCTTCTGATTAGCATAGA

4232 Right CTCATCCCCTTTGCTACATAATAGA
4225 Left ACTGCCTTTATGAAGTCATGTTAGC
4226 Right GGTATCCCGTATTACCTCAAACTCT A233 Left TTTTAACACTTTGCCAGACACTGTA
234 Right GGTATCCCGTATTACCTCAAACTCT
4227 Left AGAATCTTCCATGTTTTTCAGACAG
4228 Right CTCATCCCCTTTGCTACATAATAGA A235 Left AATACATGCATCACACCATACTGTC
236 Right CTCATCCCCTTTGCTACATAATAGA
_________________________________________ :237 Left AGAATCTTCCATGTTTTTCAGACAG
4229 Left TTTTAACACTTTGCCAGACACTGTA A238 Right CATAATAGACTGGACCAATTTGAGG
4230 Right CAGAAACTAGGTGTCCTTTTTGTGT ____ ___________________________________________ 239 Left TTTAATCCAATTTAAGGGGATGTTT

4231 Left GGATGTTTAGGCTCTGTCTACCATA A240 Right CAGAAACTAGGTGTCCTTTTTGTGT
KIT Exon17 151-200 bases eq.
Primer Sequence Primer Sequence ID ID
4241 Left GTTTTCTTTTCTCCTCCAACCTAAT L261 Left TGAATTTAAATGGTTTTCTTTTCTCC
4242 Right ACTGTCAAGCAGAGAATGGGTACT L262 Right CTGTCAAGCAGAGAATGGGTACT
4243 Left GTTTTCTTTTCTCCTCCAACCTAAT L263 Left TGAATTTAAATGGTTTTCTTTTCTCC
4244 Right CTGTCAAGCAGAGAATGGGTACT L264 Right GACTGTCAAGCAGAGAATGGGTACT
4245 Left TGAATTTAAATGGTTTTCTTTTCTCC L265 Left TTTTCTTTTCTCCTCCAACCTAATA
4246 Right ACTGTCAAGCAGAGAATGGGTACT L266 Right CTGTCAAGCAGAGAATGGGTACT
4247 Left GTTTTCTTTTCTCCTCCAACCTAAT L267 Left GTTTTCTTTTCTCCTCCAACCTAA
4248 Right GACTGTCAAGCAGAGAATGGGTACT L268 Right ACTGTCAAGCAGAGAATGGGTACT
4249 Left TTTTCTTTTCTCCTCCAACCTAATA L269 Left TTTTCTTTTCTCCTCCAACCTAATA
4250 Right ACTGTCAAGCAGAGAATGGGTACT L270 Right GACTGTCAAGCAGAGAATGGGTACT
4251 Left TAAATGGTTTTCTTTTCTCCTCCA L271 Left TAAATGGTTTTCTTTTCTCCTCCA
4252 Right ACTGTCAAGCAGAGAATGGGTACT L272 Right CTGTCAAGCAGAGAATGGGTACT
4253 Left TAAATGGTTTTCTTTTCTCCTCCAA L273 Left TAAATGGTTTTCTTTTCTCCTCCA
4254 Right ACTGTCAAGCAGAGAATGGGTACT L274 Right GACTGTCAAGCAGAGAATGGGTACT
4255 Left TTAAATGGTTTTCTTTTCTCCTCCA L275 Left GTTTTCTTTTCTCCTCCAACCTAAT
4256 Right ACTGTCAAGCAGAGAATGGGTACT L276 Right CAGGACTGTCAAGCAGAGAATG
4257 Left GTTTTCTTTTCTCCTCCAACCTAATA L277 Left TAAATGGTTTTCTTTTCTCCTCCAA
4258 Right ACTGTCAAGCAGAGAATGGGTACT L278 Right CTGTCAAGCAGAGAATGGGTACT
4259 Left GTTTTCTTTTCTCCTCCAACCTAAT L279 Left TTAAATGGTTTTCTTTTCTCCTCCA
4260 Right GACTGTCAAGCAGAGAATGGGTA L280 Right CTGTCAAGCAGAGAATGGGTACT
KIT Exon17 201-300 S eq. . z 283 Left GTTTTCTTTTCTCCTCCAACCTAAT
Pnmer Sequence ID z 284 Right AAATGTGTGATATCCCTAGACAGGA
4281 Left GTTTTCTTTTCTCCTCCAACCTAAT
4282 Right ATCACAGGAAACAATTTTTATCGAA
___________________________________________ Seq. Primer Sequence ID 4302 Right AAATGTGTGATATCCCTAGACAGGA
4285 Left GTTTTCTTTTCTCCTCCAACCTAAT
4286 Right TGTGTGATATCCCTAGACAGGATTT 4303 Left TTTTCTTTTCTCCTCCAACCTAATA
4304 Right TGTGTGATATCCCTAGACAGGATTT
4287 Left GTTTTCTTTTCTCCTCCAACCTAAT
4288 Right CACAGGAAACAATTTTTATCGAAAG 4305 Left TGAATTTAAATGGTTTTCTTTTCTCC
4306 Right CACAGGAAACAATTTTTATCGAAAG
_________________________________________ 7307 Left TGAATTTAAATGGTTTTCTTTTCTCC

4289 Left GTTTTCTTTTCTCCTCCAACCTAAT 4308 Right AAAATGTGTGATATCCCTAGACAGG
4290 Right AAAATGTGTGATATCCCTAGACAGG ____ 4309 Left TTTTCTTTTCTCCTCCAACCTAATA
4291 Left TGAATTTAAATGGTTTTCTTTTCTCC 4310 Right CACAGGAAACAATTTTTATCGAAAG
4292 Right TGTGTGATATCCCTAGACAGGATTT ____ 4311 Left TAAATGGTTTTCTTTTCTCCTCCA
4293 Left TGAATTTAAATGGTTTTCTTTTCTCC 4312 Right ATCACAGGAAACAATTTTTATCGAA
4294 Right AAATGTGTGATATCCCTAGACAGGA ______ 4313 Left TAAATGGTTTTCTTTTCTCCTCCA
4295 Left TTTTCTTTTCTCCTCCAACCTAATA 4314 Right AAATGTGTGATATCCCTAGACAGGA
4296 Right ATCACAGGAAACAATTTTTATCGAA _____ __________________________________________ 4315 Left TAAATGGTTTTCTTTTCTCCTCCA

4297 Left GTTTTCTTTTCTCCTCCAACCTAAT 4316 Right TGTGTGATATCCCTAGACAGGATTT
4298 Right ATGTGTGATATCCCTAGACAGGATT _____ __________________________________________ 4317 Left TTTTCTTTTCTCCTCCAACCTAATA
4299 Left GTTTTCTTTTCTCCTCCAACCTAAT 4318 Right AAAATGTGTGATATCCCTAGACAGG
4300 Right AATGTGTGATATCCCTAGACAGGAT ____ 4319 Left TAAATGGTTTTCTTTTCTCCTCCAA
4301 Left TTTTCTTTTCTCCTCCAACCTAATA 4320 Right ATCACAGGAAACAATTTTTATCGAA
KIT Exon17 301-400 Seq. . z 335 Left TTCAAGGCGTACTTTTGATTTTTAT
Pnmer Sequence ID z 336 Right TCGAAAGTTGAAACTAAAAATCCTTT
4321 Le ft ATTCAAGGCGTACTTTTGATTTTTA
4322 Right ATCACAGGAAACAATTTTTATCGAA z 337 Left ATTCAAGGCGTACTTTTGATTTTTA
z 338 Right TCACAGGAAACAATTTTTATCGAA
4323 Left TCAAGGCGTACTTTTGATTTTTATT Seq. .
S
4324 Right ATCACAGGAAACAATTTTTATCGAA ID Primer equence z 339 Left TCAAGGCGTACTTTTGATTTTTATT
4325 Le ft TTCAAGGCGTACTTTTGATTTTTAT z 340 Right TCACAGGAAACAATTTTTATCGAA
4326 Right ATCACAGGAAACAATTTTTATCGAA
z 341 Left TTCAAGGCGTACTTTTGATTTTTAT
4327 Le ft ATTCAAGGCGTACTTTTGATTTTTA z 342 Right TCACAGGAAACAATTTTTATCGAA
4328 Right CACAGGAAACAATTTTTATCGAAAG
z 343 Left ATTCAAGGCGTACTTTTGATTTTT
4329 Le ft GTTTTCTTTTCTCCTCCAACCTAAT z 344 Right ATCACAGGAAACAATTTTTATCGAA
4330 Right TAGTAATGTTCAGCATACCATGCAA
z 345 Left TTCAAGGCGTACTTTTGATTTTTATT
4331 Left ATTCAAGGCGTACTTTTGATTTTTAT z 346 Right ATCACAGGAAACAATTTTTATCGAA
4332 Right ATCACAGGAAACAATTTTTATCGAA
z 347 Left AGTCCTGAGAAGAAAACAGCATTTA
4333 Le ft ATCATTCAAGGCGTACTTTTGATTT z 348 Right ACTGTCAAGCAGAGAATGGGTACT
4334 Right ATCACAGGAAACAATTTTTATCGAA
z 349 Left ATTCAAGGCGTACTTTTGATTTTT

4350 Right CACAGGAAACAATTTTTATCGAAAG z 355 Left TTCAAGGCGTACTTTTGATTTTTA
z 356 Right ATCACAGGAAACAATTTTTATCGAA
4351 Left GTTTTCTTTTCTCCTCCAACCTAAT
4352 Right GTAATGTTCAGCATACCATGCAAAT z 357 Left GAACATCATTCAAGGCGTACTTTT
z 358 Right ATCACAGGAAACAATTTTTATCGAA
4353 Left CATTCAAGGCGTACTTTTGATTTT
4354 Right ATCACAGGAAACAATTTTTATCGAA z 359 Left CATTCAAGGCGTACTTTTGATTTTT
z 360 Right ATCACAGGAAACAATTTTTATCGAA
KIT Exon17 401-500 bases Seq.
Primer Sequence Primer Sequence ID ID
4361 Left ATGTATTTCCCTATGAATGAAAGCA L381 Left GTTTTCTTTTCTCCTCCAACCTAAT
4362 Right ATCACAGGAAACAATTTTTATCGAA L382 Right AATCAAGTTCATTGCTATTCTCAGG
4363 Left TATTTCCCTATGAATGAAAGCAGTC L383 Left TCCTGAGAAGAAAACAGCATTTATT
4364 Right ATCACAGGAAACAATTTTTATCGAA L384 Right CACAGGAAACAATTTTTATCGAAAG
4365 Left AGTCCTGAGAAGAAAACAGCATTTA L385 Left GTTTTCTTTTCTCCTCCAACCTAAT
4366 Right ATCACAGGAAACAATTTTTATCGAA L386 Right GCAAAATCAAGTTCATTGCTATTCT
4367 Left AGTCCTGAGAAGAAAACAGCATTTA L387 Left GTTTTCTTTTCTCCTCCAACCTAAT
4368 Right TGTGTGATATCCCTAGACAGGATTT L388 Right AGCAAAATCAAGTTCATTGCTATTC
4369 Left ATGTATTTCCCTATGAATGAAAGCA L389 Left TCCTGAGAAGAAAACAGCATTTATT
4370 Right CACAGGAAACAATTTTTATCGAAAG L390 Right AAAATGTGTGATATCCCTAGACAGG
4371 Left TATTTCCCTATGAATGAAAGCAGTC L391 Left TCAAGGCGTACTTTTGATTTTTATT
4372 Right CACAGGAAACAATTTTTATCGAAAG L392 Right AAATGTGTGATATCCCTAGACAGGA
4373 Left AGTCCTGAGAAGAAAACAGCATTTA L393 Left TCAAGGCGTACTTTTGATTTTTATT
4374 Right CACAGGAAACAATTTTTATCGAAAG L394 Right TGTGTGATATCCCTAGACAGGATTT
4375 Left TCCTGAGAAGAAAACAGCATTTATT L395 Left TTCAAGGCGTACTTTTGATTTTTAT
4376 Right ATCACAGGAAACAATTTTTATCGAA L396 Right AAATGTGTGATATCCCTAGACAGGA
4377 Left TCCTGAGAAGAAAACAGCATTTATT L397 Left ATTCAAGGCGTACTTTTGATTTTTA
4378 Right TGTGTGATATCCCTAGACAGGATTT L398 Right AAATGTGTGATATCCCTAGACAGGA
4379 Left AGTCCTGAGAAGAAAACAGCATTTA L399 Left TTCAAGGCGTACTTTTGATTTTTAT
4380 Right AAAATGTGTGATATCCCTAGACAGG L400 Right TGTGTGATATCCCTAGACAGGATTT
KIT Exon17 501-600 S eq. .
Primer S equence ID L407 Left TATTTCCCTATGAATGAAAGCAGTC

4401 Left ATGTATTTCCCTATGAATGAAAGCA z 408 Right TGTGTGATATCCCTAGACAGGATTT
4402 Right AAATGTGTGATATCCCTAGACAGGA
L409 Left GATTGAATTTGCAAAGGCATATTAG
4403 Left ATGTATTTCCCTATGAATGAAAGCA z 410 Right ATCACAGGAAACAATTTTTATCGAA
4404 Right TGTGTGATATCCCTAGACAGGATTT Seq. .
ID Primer Sequence 4405 Left TATTTCCCTATGAATGAAAGCAGTC z 411 Left ATGTATTTCCCTATGAATGAAAGCA
4406 Right AAATGTGTGATATCCCTAGACAGGA L412 Right AAAATGTGTGATATCCCTAGACAGG

4413 Le ft TATTTCCCTATGAATGAAAGCAGTC z 427 Left TCAAGGCGTACTTTTGATTTTTATT
4414 Right AAAATGTGTGATATCCCTAGACAGG z 428 Right GCAAAATCAAGTTCATTGCTATTCT
4415 Left GATTGAATTTGCAAAGGCATATTAG z 429 Left TTCAAGGCGTACTTTTGATTTTTAT
4416 Right CACAGGAAACAATTTTTATCGAAAG L430 Right GCAAAATCAAGTTCATTGCTATTCT
4417 Left AAAGGCATATTAGGAACTCTGTGAA z 431 Left ATTCAAGGCGTACTTTTGATTTTTA
4418 Right ATCACAGGAAACAATTTTTATCGAA L432 Right GCAAAATCAAGTTCATTGCTATTCT
4419 Left AAGGCATATTAGGAACTCTGTGAAA L433 Left TCAAGGCGTACTTTTGATTTTTATT
4420 Right ATCACAGGAAACAATTTTTATCGAA z 434 Right AGCAAAATCAAGTTCATTGCTATTC
4421 Le ft TCAAGGCGTACTTTTGATTTTTATT L435 Left TTCAAGGCGTACTTTTGATTTTTAT
4422 Right AATCAAGTTCATTGCTATTCTCAGG z 436 Right AGCAAAATCAAGTTCATTGCTATTC
4423 Le ft TTCAAGGCGTACTTTTGATTTTTAT z 437 Left ATTCAAGGCGTACTTTTGATTTTTA
4424 Right AATCAAGTTCATTGCTATTCTCAGG z 438 Right AGCAAAATCAAGTTCATTGCTATTC
4425 Le ft ATTCAAGGCGTACTTTTGATTTTTA z 439 Left AAAGGCATATTAGGAACTCTGTGAA
4426 Right AATCAAGTTCATTGCTATTCTCAGG z 440 Right TGTGTGATATCCCTAGACAGGATTT
KIT Exon17 601-800 S eq.
Primer Sequence ID Seq. .
S
4441 Left TATTTCCCTATGAATGAAAGCAGTC ID Primer equence 4442 Right AATCAAGTTCATTGCTATTCTCAGG z 461 Left GATTGAATTTGCAAAGGCATATTAG
z 462 Right GCAAAATCAAGTTCATTGCTATTCT
4443 Le ft TATTTCCCTATGAATGAAAGCAGTC
4444 Right GCAAAATCAAGTTCATTGCTATTCT z 463 Left GATTGAATTTGCAAAGGCATATTAG
z 464 Right AGCAAAATCAAGTTCATTGCTATTC
4445 Le ft ACATTTCCCAACAATTACCAAACTA
4446 Right ATCACAGGAAACAATTTTTATCGAA z 465 Left TTACCAGTCCTACCCTTAAATGTCA
z 466 Right ATCACAGGAAACAATTTTTATCGAA
4447 Le ft ACATTTCCCAACAATTACCAAACTA
4448 Right AAATGTGTGATATCCCTAGACAGGA z 467 Left TCCCAACAATTACCAAACTAAGAAA
z 468 Right AAATGTGTGATATCCCTAGACAGGA
4449 Le ft ACATTTCCCAACAATTACCAAACTA
4450 Right TGTGTGATATCCCTAGACAGGATTT z 469 Left TTCCCAACAATTACCAAACTAAGAA
z 470 Right AAATGTGTGATATCCCTAGACAGGA
4451 Left GAAGGTTAGGAATGGAAAGAATGAT
4452 Right ATCACAGGAAACAATTTTTATCGAA z 471 Left TTTCCCAACAATTACCAAACTAAGA
z 472 Right AAATGTGTGATATCCCTAGACAGGA
4453 Le ft GATTGAATTTGCAAAGGCATATTAG
4454 Right AATCAAGTTCATTGCTATTCTCAGG z 473 Left TTCCCAACAATTACCAAACTAAGAA
z 474 Right TGTGTGATATCCCTAGACAGGATTT
4455 Le ft TTCCCAACAATTACCAAACTAAGAA
4456 Right ATCACAGGAAACAATTTTTATCGAA z 475 Left TTTCCCAACAATTACCAAACTAAGA
z 476 Right TGTGTGATATCCCTAGACAGGATTT
4457 Le ft TCCCAACAATTACCAAACTAAGAAA
4458 Right ATCACAGGAAACAATTTTTATCGAA z 477 Left TCCCAACAATTACCAAACTAAGAAA
z 478 Right TGTGTGATATCCCTAGACAGGATTT
4459 Le ft TTTCCCAACAATTACCAAACTAAGA
4460 Right ATCACAGGAAACAATTTTTATCGAA L479 Left ATTAACCGAACAGAATGAGTTACCA

4480 Right ATCACAGGAAACAATTTTTATCGAA
KIT Exon17 801-1000 bases S eq. Primer Sequence ID 4501 Left TTTCCCAACAATTACCAAACTAAGA
4481 Left ACATTTCCCAACAATTACCAAACTA 4502 Right GCAAAATCAAGTTCATTGCTATTCT
4482 Right AATCAAGTTCATTGCTATTCTCAGG
S eq. . mer 4503 Left TTTCCCAACAATTACCAAACTAAGA
Pri Sequence ID 4504 Right AGCAAAATCAAGTTCATTGCTATTC
4483 Left ACATTTCCCAACAATTACCAAACTA ______ 4484 Right GCAAAATCAAGTTCATTGCTATTCT 4505 Left GGGTGAAGCATAGACTTGAGTTTTA
4485 Left ACATTTCCCAACAATTACCAAACTA 4506 Right ATCACAGGAAACAATTTTTATCGAA
4486 Right AGCAAAATCAAGTTCATTGCTATTC
4507 Left ATTAACCGAACAGAATGAGTTAC CA
4487 Left ATGTATTTCCCTATGAATGAAAGCA 4508 Right AATCAAGTTCATTGCTATTCTCAGG
4488 Right CATGCCTTAGTTTCTCCAACTTTTA
4509 Left TTACCAGTCCTACCCTTAAATGTCA
4489 Left TATTTCCCTATGAATGAAAGCAGTC 4510 Right GCAAAATCAAGTTCATTGCTATTCT
4490 Right CATGCCTTAGTTTCTCCAACTTTTA
4511 Left TTACCAGTCCTACCCTTAAATGTCA
4491 Left TCCCAACAATTACCAAACTAAGAAA 4512 Right AGCAAAATCAAGTTCATTGCTATTC
4492 Right AATCAAGTTCATTGCTATTCTCAGG _____ 4513 Left GGGTGAAGCATAGACTTGAGTTTTA
4493 Left TTTCCCAACAATTACCAAACTAAGA 4514 Right AAATGTGTGATATCCCTAGACAGGA
4494 Right AATCAAGTTCATTGCTATTCTCAGG _____ 4515 Left GGGTGAAGCATAGACTTGAGTTTTA
4495 Left TCCCAACAATTACCAAACTAAGAAA 4516 Right TGTGTGATATCCCTAGACAGGATTT
4496 Right GCAAAATCAAGTTCATTGCTATTCT _____ 4517 Left AGTCCTGAGAAGAAAACAGCATTTA
4497 Left TCCCAACAATTACCAAACTAAGAAA 4518 Right CATGCCTTAGTTTCTCCAACTTTTA
4498 Right AGCAAAATCAAGTTCATTGCTATTC
4519 Left ATTAACCGAACAGAATGAGTTAC CA
4499 Left TTCCCAACAATTACCAAACTAAGAA 4520 Right GCAAAATCAAGTTCATTGCTATTCT
4500 Right AGCAAAATCAAGTTCATTGCTATTC
KIT Exon17 2kb S eq. . z 532 Right AGTATGTAACCACCGTCACGATTAT
ID Pnmer Sequence 4521 Left AACCAAAAGCAGAGGAAATTTAGTT L533 Left GGGTGAAGCATAGACTTGAGTTTTA
4522 Right AACACTCTTACAAAACCAAATCGAG z 534 Right TTGGACAACTCTTGACAAAATTACA
4523 Left AGCCATAGTTAAAATGCAGAATGTC L535 Left GAAGGTTAGGAATGGAAAGAATGAT
4524 Right AACACTCTTACAAAACCAAATCGAG z 536 Right AGTATGTAACCACCGTCACGATTAT
__________________________________________ Seq.
Primer Sequence 4525 Left GAGCCATAGTTAAAATGCAGAATGT ID
4526 Right AACACTCTTACAAAACCAAATCGAG L537 Left GGGTGAAGCATAGACTTGAGTTTTA
z 538 Right ATTCAGAGGTATTGGACAACTCTTG
4527 Left AGAGCCATAGTTAAAATGCAGAATG
4528 Right AACACTCTTACAAAACCAAATCGAG L539 Left GAAGGTTAGGAATGGAAAGAATGAT
z 540 Right TTGGACAACTCTTGACAAAATTACA
4529 Left ACATTTCCCAACAATTACCAAACTA
4530 Right AGTATGTAACCACCGTCACGATTAT z 541 Left ATGTATTTCCCTATGAATGAAAGCA
z 542 Right AAAACCCACAATTACTTTTACACCA
4531 Left GGGTGAAGCATAGACTTGAGTTTTA

4543 Left TATTTCCCTATGAATGAAAGCAGTC z 552 Right GGCAGAGAATATTATAAAGGGCAAT
4544 Right AAAACCCACAATTACTTTTACACCA
z 553 Left AGAGCCATAGTTAAAATGCAGAATG
4545 Left AACCAAAAGCAGAGGAAATTTAGTT z 554 Right GGCAGAGAATATTATAAAGGGCAAT
4546 Right ATTACATTATCATAAGGGGCACAAA
z 555 Left AGCCATAGTTAAAATGCAGAATGTC
4547 Left GAAGGTTAGGAATGGAAAGAATGAT z 556 Right ATTACATTATCATAAGGGGCACAAA
4548 Right ATTCAGAGGTATTGGACAACTCTTG
z 557 Left GAGCCATAGTTAAAATGCAGAATGT
4549 Le ft AGCCATAGTTAAAATGCAGAATGTC L558 Right ATTACATTATCATAAGGGGCACAAA
4550 Right GGCAGAGAATATTATAAAGGGCAAT
z 559 Left AGAGCCATAGTTAAAATGCAGAATG
4551 Left GAGCCATAGTTAAAATGCAGAATGT L560 Right ATTACATTATCATAAGGGGCACAAA
KIT Exon17 5kb Seq.
Primer Sequence Primer Sequence ID ID
4561 Left ATGGTTCAGAAAATCATCCAAATTA z 581 Left GTACCTACCTATCAAGCAACCAAGA
4562 Right TCTGCCATAAAAAGCTAAATCAATC z 582 Right GGCCACTAAGTTGTAAGTGCTGTAT
4563 Le ft TCAGAAAATCATCCAAATTATTGGT z 583 Left GAGTACCTACCTATCAAGCAAC CAA
4564 Right TCTGCCATAAAAAGCTAAATCAATC z 584 Right GGCCACTAAGTTGTAAGTGCTGTAT
4565 Left GTATATTGCTGCAGTTGTGTGGTAG 7 585 Left TCATGGTGTTTCTATGCTAATCAGA
4566 Right TAAGGGCTCCTAACCTGAGATCTAT z 586 Right TAAGGGCTCCTAACCTGAGATCTAT
4567 Le ft TCATGGTGTTTCTATGCTAATCAGA z 587 Left GTAACCCAGCCTAGGATTGTTAAAT
4568 Right TCTGCCATAAAAAGCTAAATCAATC z 588 Right CTTGAGTACCATCTCACAAAAACCT
4569 Le ft GTACCTACCTATCAAGCAACCAAGA z 589 Left TCATGGTGTTTCTATGCTAATCAGA
_ 4570 Right CTTGAGTACCATCTCACAAAAACCT z 590 Right GGCCACTAAGTTGTAAGTGCTGTAT
4571 Left GAGTACCTACCTATCAAGCAACCAA L591 Left AACCAAAAGCAGAGGAAATTTAGTT
4572 Right CTTGAGTACCATCTCACAAAAACCT z 592 Right CAGTGTGTCATAAAGAATCCAAGTG
_ 4573 Left TCTATGCTAATCAGAAGCAGGAAGT z 593 Left TCTATGCTAATCAGAAGCAGGAAGT
4574 Right TCTGCCATAAAAAGCTAAATCAATC z 594 Right TAAGGGCTCCTAACCTGAGATCTAT
4575 Le ft ATGGTTCAGAAAATCATCCAAATTA z 595 Left ATGTTTTTGTGCCTGAGTATCTTTC
4576 Right GGCCACTAAGTTGTAAGTGCTGTAT z 596 Right CAGTGTGTCATAAAGAATCCAAGTG
4577 Le ft TCAGAAAATCATCCAAATTATTGGT z 597 Left TCTATGCTAATCAGAAGCAGGAAGT
4578 Right GGCCACTAAGTTGTAAGTGCTGTAT z 598 Right GGCCACTAAGTTGTAAGTGCTGTAT
4579 Le ft GTATATTGCTGCAGTTGTGTGGTAG z 599 Left AACCAAAAGCAGAGGAAATTTAGTT
4580 Right AAAACCCACAATTACTTTTACACCA z 600 Right CAATTTGCAACCTAAGATTAGGAGA
Table 10. KRAS Capture Primer List for NGS Panel KRAS Exonl 169-300 bases Seq.z 601 Left CTCGGAGCTCGATTTTCCTA
Primer Sequence ID z 602 Right GGGACCCCTAATTCATTCACTC

z 622 Right GGGACCCCTAATTCATTCACTC
4603 Left CTCGGAGCTCGATTTTCCTA
4604 Right GGGACCCCTAATTCATTCACT z 623 Left CTCGGAGCTCGATTTTCCT
z 624 Right GGGACCCCTAATTCATTCACT
4605 Left CTCGGAGCTCGATTTTCCTA
4606 Right GGACCCCTAATTCATTCACTCG z 625 Left CTCGGAGCTCGATTTTCCTA
Sea z 626 Right GGGGACCCCTAATTCATTCA
ID' Primer Sequence _ _______________________________________________________________________________ ______ 4607 Left CTCGGAGCTCGATTTTCCT z 627 Left GTGCTCGGAGCTCGATTTT
4608 Right GGGACCCCTAATTCATTCACTC z 628 Right GGGACCCCTAATTCATTCACT
_ 4609 Left GTGCTCGGAGCTCGATTTT z 629 Left TCGGAGCTCGATTTTCCTA
4610 Right GGGACCCCTAATTCATTCACTC z 630 Right GGGACCCCTAATTCATTCACTC
4611 Left CTCGGAGCTCGATTTTCCTA z 631 Left CTCGGAGCTCGATTTTCCT
4612 Right GGGGACCCCTAATTCATTCACT z 632 Right GGACCCCTAATTCATTCACTCG
4613 Left GCTCGGAGCTCGATTTTCCTA z 633 Left GTGCTCGGAGCTCGATTTT
4614 Right GGGACCCCTAATTCATTCACTC z 634 Right GGACCCCTAATTCATTCACTCG
4615 Left CTCGGAGCTCGATTTTCCTA z 635 Left GTGTGCTCGGAGCTCGATT
4616 Right GACCCCTAATTCATTCACTCG z 636 Right GGGACCCCTAATTCATTCACTC
4617 Left CTCGGAGCTCGATTTTCCTA z 637 Left CTCGGAGCTCGATTTTCCTA
4618 Right GGGGACCCCTAATTCATTCAC z 638 Right GGGGACCCCTAATTCATTCACTC

_________________________________________ ¨
__________________________________________ 4619 Left GTGCTCGGAGCTCGATTTTC z 639 Left TGCTCGGAGCTCGATTTTC
4620 Right GGGACCCCTAATTCATTCACTC z 640 Right GGGACCCCTAATTCATTCACTC
4621 Left TGCTCGGAGCTCGATTTT
KRAS Exonl 301-400 bases Sea z 654 Right GGGACCCCTAATTCATTCACT
ID' Primer Sequence 4641 Left CCCGTCTGAAGAAGAATCGAG z 655 Left TACGCCCGTCTGAAGAAGAATC
4642 Right GGGACCCCTAATTCATTCACTC z 656 Right GGGACCCCTAATTCATTCACTC
_ 4643 Left GTACGCCCGTCTGAAGAAGA z 657 Left GTACGCCCGTCTGAAGAAGAAT
4644 Right GGGACCCCTAATTCATTCACTC z 658 Right GGACCCCTAATTCATTCACTCG
4645 Left CATCGATAGCTCTGCCCTCT z 659 Left TACGCCCGTCTGAAGAAGAA
4646 Right GGGACCCCTAATTCATTCACTC z 660 Right GGGACCCCTAATTCATTCACTC
_________________________________________ Seq.
4647 Left ATCGATAGCTCTGCCCTCTG ID Primer Sequence 4648 Right GGGACCCCTAATTCATTCACTC z 661 Left GGAACGCATCGATAGCTCTG
z 662 Right GGGACCCCTAATTCATTCACTC
4649 Left GTACGCCCGTCTGAAGAAGAATC
4650 Right GGGACCCCTAATTCATTCACTC z 663 Left TACGCCCGTCTGAAGAAGAAT
z 664 Right GGGACCCCTAATTCATTCACT
4651 Left GTACGCCCGTCTGAAGAAGAA
4652 Right GGGACCCCTAATTCATTCACTC z 665 Left TACGCCCGTCTGAAGAAGAAT
z 666 Right GGACCCCTAATTCATTCACTCG
4653 Left GTACGCCCGTCTGAAGAAGAAT

4667 Left CCCGTCTGAAGAAGAATCGAG z 674 Right GGGGACCCCTAATTCATTCACT
4668 Right GGGACCCCTAATTCATTCACT
z 675 Left ATCGATAGCTCTGCCCTCTG
4669 Left CATCGATAGCTCTGCCCTCTG z 676 Right GGGACCCCTAATTCATTCACT
4670 Right GGGACCCCTAATTCATTCACTC
z 677 Left CATCGATAGCTCTGCCCTCT
4671 Left GTACGCCCGTCTGAAGAAGA z 678 Right GGGACCCCTAATTCATTCACT
4672 Right GGGACCCCTAATTCATTCACT
z 679 Left GTACGCCCGTCTGAAGAAGAATC
4673 Left GTACGCCCGTCTGAAGAAGAAT z 680 Right GGGACCCCTAATTCATTCACT
KRAS Exonl 401-600 bases Sea Seq.
Primer Sequence Primer Sequence ID
4681 Left CTCTTCCCTCTTCCCACACC L701 Left CCTCTTCCCTCTTCCCACA
4682 Right GGGACCCCTAATTCATTCACTC L702 Right GGGACCCCTAATTCATTCACT
4683 Left CCTCTTCCCTCTTCCCACAC L703 Left CCTCTTCCCTCTTCCCACA
4684 Right GGGACCCCTAATTCATTCACTC L704 Right GGACCCCTAATTCATTCACTCG
4685 Left TCTTCCCTCTTCCCACACC L705 Left GTACGCCCGTCTGAAGAAGAAT
4686 Right GGGACCCCTAATTCATTCACTC L706 Right GAGGAGGAAGGAAGGGGTTC
4687 Left CCTCTTCCCTCTTCCCACA L707 Left GTACGCCCGTCTGAAGAAGAAT
4688 Right GGGACCCCTAATTCATTCACTC L708 Right AGGAGGAAGGAAGGGGTTC
4689 Left CTCTTCCCTCTTCCCACACC L709 Left TACGCCCGTCTGAAGAAGAAT
4690 Right GGGACCCCTAATTCATTCACT L710 Right GAGGAGGAAGGAAGGGGTTC
4691 Left CCTCTTCCCTCTTCCCACAC L711 Left CTCTTCCCTCTTCCCACACC
4692 Right GGGACCCCTAATTCATTCACT L712 Right GGGGACCCCTAATTCATTCACT
4693 Left TCTTCCCTCTTCCCACACC L713 Left CCTCTTCCCTCTTCCCACAC
4694 Right GGGACCCCTAATTCATTCACT L714 Right GGGGACCCCTAATTCATTCACT
4695 Left CTCTTCCCTCTTCCCACACC L715 Left TCTTCCCTCTTCCCACACC
4696 Right GGACCCCTAATTCATTCACTCG L716 Right GGGGACCCCTAATTCATTCACT
4697 Left CCTCTTCCCTCTTCCCACAC L717 Left CTCTTCCCTCTTCCCACACC
4698 Right GGACCCCTAATTCATTCACTCG L7 18 Right GACCCCTAATTCATTCACTCG
4699 Left TCTTCCCTCTTCCCACACC L719 Left CCTCTTCCCTCTTCCCACAC
4700 Right GGACCCCTAATTCATTCACTCG L720 Right GACCCCTAATTCATTCACTCG
KRAS Exonl 601-800 bases Seq.
Primer Sequence ID z 725 Left TACGCCCGTCTGAAGAAGAAT
4721 Left GTACGCCCGTCTGAAGAAGAAT z 726 Right GTTTATACCTTCGTCCTAGAGATGC
4722 Right GTTTATACCTTCGTCCTAGAGATGC
z 727 Left CCCGTCTGAAGAAGAATCGAG
4723 Left GTACGCCCGTCTGAAGAAGAAT z 728 Right GTTTATACCTTCGTCCTAGAGATGC
4724 Right AGTTTATACCTTCGTCCTAGAGATGC

4729 Left TACGCCCGTCTGAAGAAGAAT 4743 Left CTCGGAGCTCGATTTTCCTA
4730 Right AGTTTATACCTTCGTCCTAGAGATGC 4744 Right GTTTATACCTTCGTCCTAGAGATGC
__________________________________________ 4745 Left CCCGTCTGAAGAAGAATCGAG
Sea 7746 Right GGAGAAGTTTATACCTTCGTCCTAGA
ID' Primer Sequence 4731 Left GTACGCCCGTCTGAAGAAGA 4747 Left GTACGCCCGTCTGAAGAAGA
4732 Right GTTTATACCTTCGTCCTAGAGATGC 4748 Right AGTTTATACCTTCGTCCTAGAGATGC
4733 Left ATCGATAGCTCTGCCCTCTG 4749 Left ATCGATAGCTCTGCCCTCTG
4734 Right GTTTATACCTTCGTCCTAGAGATGC 4750 Right AGTTTATACCTTCGTCCTAGAGATGC
4735 Left CATCGATAGCTCTGCCCTCT 4751 Left CATCGATAGCTCTGCCCTCT
4736 Right GTTTATACCTTCGTCCTAGAGATGC 4752 Right AGTTTATACCTTCGTCCTAGAGATGC
4737 Left GTACGCCCGTCTGAAGAAGAATC 4753 Left GTACGCCCGTCTGAAGAAGAATC
4738 Right GTTTATACCTTCGTCCTAGAGATGC 4754 Right AGTTTATACCTTCGTCCTAGAGATGC
4739 Left CCCGTCTGAAGAAGAATCGAG 4755 Left CTCGGAGCTCGATTTTCCTA
4740 Right AGTTTATACCTTCGTCCTAGAGATGC 4756 Right AGTTTATACCTTCGTCCTAGAGATGC
4757 Left ATCGATAGCTCTGCCCTCTG
4758 Right GGAGAAGTTTATACCTTCGTCCTAGA
4741 Left CTCGGAGCTCGATTTTCCTA
4742 Right CTTACTCACACATCCCCTACACAC 4759 Left CATCGATAGCTCTGCCCTCT
4760 Right GGAGAAGTTTATACCTTCGTCCTAGA
KRAS Exonl 801-1000 bases Seq. z 777 Left TACGCCCGTCTGAAGAAGAAT
Primer Sequence ID z 778 Right ACAGTGGTCTCTAAGCACTTTCCTA
4761 Left ACTTTTGGTGACTGCTTGTTTATTT
4762 Right GGGACCCCTAATTCATTCACTC z 779 Left TACGCCCGTCTGAAGAAGAAT
z 780 Right AAACACAATAACCTCAAACAGTGGT
4763 Left CACTTTTGGTGACTGCTTGTTTATT
4764 Right GGGACCCCTAATTCATTCACTC Seq.
ID Primer Sequence 4765 Left GTACGCCCGTCTGAAGAAGAAT z 781 Left CACTTTTGGTGACTGCTTGTTTAT
4766 Right ACAGTGGTCTCTAAGCACTTTCCTA z 782 Right GGGACCCCTAATTCATTCACTC
4767 Left GTACGCCCGTCTGAAGAAGAAT z 783 Left ACTTTTGGTGACTGCTTGTTTATTT
4768 Right AAACACAATAACCTCAAACAGTGGT z 784 Right GGGACCCCTAATTCATTCACT
4769 Left CTTTTGGTGACTGCTTGTTTATTTA z 785 Left GTACGCCCGTCTGAAGAAGAAT
4770 Right GGGACCCCTAATTCATTCACTC z 786 Right ACAATAACCTCAAACAGTGGTCTCT
4771 Left GTACGCCCGTCTGAAGAAGAAT z 787 Left CTTTTGGTGACTGCTTGTTTATTTAC
4772 Right TAACCTCAAACAGTGGTCTCTAAGC z 788 Right GGGACCCCTAATTCATTCACTC
4773 Left GTACGCCCGTCTGAAGAAGAAT z 789 Left ACTTTTGGTGACTGCTTGTTTATTTA
4774 Right ACACAATAACCTCAAACAGTGGTCT z 790 Right GGGACCCCTAATTCATTCACTC
4775 Left TTTTGGTGACTGCTTGTTTATTTAC z 791 Left ACTTTTGGTGACTGCTTGTTTATTT
4776 Right GGGACCCCTAATTCATTCACTC z 792 Right GGACCCCTAATTCATTCACTCG

4793 Left TACGCCCGTCTGAAGAAGAAT z 797 Left TACGCCCGTCTGAAGAAGAAT
4794 Right TAACCTCAAACAGTGGTCTCTAAGC z 798 Right ACACAATAACCTCAAACAGTGGTCT
4795 Left CCCGTCTGAAGAAGAATCGAG z 799 Left CCCGTCTGAAGAAGAATCGAG
4796 Right ACAGTGGTCTCTAAGCACTTTCCTA z 800 Right AAACACAATAACCTCAAACAGTGGT
KRAS Exonl 2kb S eq. Primer Sequence ID 4821 Left GGGATTTAAATTCAGCTTTATTGGT

4801 Left GGGATTTAAATTCAGCTTTATTGGT 4822 Right TAACCTCAAACAGTGGTCTCTAAGC
4802 Right ACAGTGGTCTCTAAGCACTTTCCTA
S eq. . 4823 Left TCAAGACTCTCCCAAGATACATTTC

Primer Sequence ID 4824 Right ATAAGAAATAGGGGAAAGGACAAGA
4803 Left GGGATTTAAATTCAGCTTTATTGGT _____ 4804 Right CATCTGGGATTAACTTTTTCCTTTT 4825 Left TCAAGACTCTCCCAAGATACATTTC
4805 Left TCAAGACTCTCCCAAGATACATTTC 4826 Right TAACCTCAAACAGTGGTCTCTAAGC
4806 Right TTTGCTATTGCTGTCTACACTCAAC
4827 Left TTCAGCTTTATTGGTGGTTTATGAT
4807 Left TCAAGACTCTCCCAAGATACATTTC 4828 Right ACAGTGGTCTCTAAGCACTTTCCTA
4808 Right ACAGTGGTCTCTAAGCACTTTCCTA
4829 Left ATTCAGCTTTATTGGTGGTTTATGA
4809 Left GGGATTTAAATTCAGCTTTATTGGT 4830 Right ACAGTGGTCTCTAAGCACTTTCCTA
4810 Right AAACACAATAACCTCAAACAGTGGT
4831 Left GGGATTTAAATTCAGCTTTATTGGT
4811 Left TCAAGACTCTCCCAAGATACATTTC 4832 Right ACACAATAACCTCAAACAGTGGTCT
4812 Right CATCTGGGATTAACTTTTTCCTTTT ____ 4833 Left TTCAGCTTTATTGGTGGTTTATGAT
4813 Left TCAAGACTCTCCCAAGATACATTTC 4834 Right CATCTGGGATTAACTTTTTCCTTTT
4814 Right AAACACAATAACCTCAAACAGTGGT _____ 4835 Left ATTCAGCTTTATTGGTGGTTTATGA
4815 Left GTAGAAAGGAAAGGATGACAGTTGA 4836 Right CATCTGGGATTAACTTTTTCCTTTT
4816 Right AAACACAATAACCTCAAACAGTGGT _____ 4837 Left ATTCAGCTTTATTGGTGGTTTATGA
4817 Left GATTACAGCCCGTGTAAGAGTAGAA 4838 Right AAACACAATAACCTCAAACAGTGGT
4818 Right ACAGTGGTCTCTAAGCACTTTCCTA
4839 Left TTCAGCTTTATTGGTGGTTTATGAT
4819 Left GATTACAGCCCGTGTAAGAGTAGAA 4840 Right AAACACAATAACCTCAAACAGTGGT
4820 Right AAACACAATAACCTCAAACAGTGGT
KRAS Exonl 5kb S eq. .
Primer S equence ID z 849 Left TGCTTTGAATGTTAGTCACAGAGAG
4841 Left GGGATTTAAATTCAGCTTTATTGGT L850 Right TGATGGATCTCAAGATTTAAGAAGG
4842 Right CAAAGCAATTAGGAATAGATGAGGA
z 851 Left ACGTAAGTAAGGAAGGGAGAACAGT
4843 Left ACGTAAGTAAGGAAGGGAGAACAGT z 852 Right AACAGTTCTCAAAATGTGGTCTAGG
4844 Right CAAAGCAATTAGGAATAGATGAGGA
z 853 Left ACGTAAGTAAGGAAGGGAGAACAGT
4845 Left AGCAGTAAATGAAACAGACCAAAAC z 854 Right TCCTTTCCCTCATGTAACACATAAT
4846 Right CAAAGCAATTAGGAATAGATGAGGA_ ________________________________________________ z 855 Left CGAATCATGAGCCTAGATGATAACT
4847 Left GGGATTTAAATTCAGCTTTATTGGT z 856 Right ATCCAACAATTTTGTAATGGAAGAA
4848 Right TCCTTTCCCTCATGTAACACATAAT Seq. Primer Sequence ID
4857 Left CGAATCATGAGCCTAGATGATAACT z 869 Left CTCCTTTTAAAAACACTTTGGAACA
4858 Right AATCCAACAATTTTGTAATGGAAGA z 870 Right CAAAGCAATTAGGAATAGATGAGGA
4859 Left CGAATCATGAGCCTAGATGATAACT L871 Left ACGTAAGTAAGGAAGGGAGAACAGT
4860 Right GAATCCAACAATTTTGTAATGGAAG z 872 Right TCATGTTACCAAGTAATGGGCTTAT
4861 Left AGCAGTAAATGAAACAGACCAAAAC L873 Left GAAGAGGGTAGGGGATATCAAATAA
4862 Right AACAGTTCTCAAAATGTGGTCTAGG z 874 Right CAAAGCAATTAGGAATAGATGAGGA
4863 Left AGCTCTGGAGAAAAAGTAGGAAAAG z 875 Left GCTGGAAATTTAGCAGTAAATGAAA
4864 Right CAAAGCAATTAGGAATAGATGAGGA z 876 Right CAAAGCAATTAGGAATAGATGAGGA
4865 Left TCAAGACTCTCCCAAGATACATTTC z 877 Left GGAAACAAGAACTTATCATGCACTT
4866 Right TCCTTTCCCTCATGTAACACATAAT z 878 Right ATCCAACAATTTTGTAATGGAAGAA
4867 Left TAGAGTGACTATGATCCGACATGAA z 879 Left GGAAACAAGAACTTATCATGCACTT
4868 Right CAAAGCAATTAGGAATAGATGAGGA z 880 Right GAATCCAACAATTTTGTAATGGAAG
KRAS Exon2 169-210 bases Seq. .
ACATGTTCTAATATAGTCACATTTTC
Primer Sequence 4899 Left ID AT
4881 Left CATGTTCTAATATAGTCACATTTTCA 4900 Right AGAATGGTCCTGCACCAGTAATA
4882 Right AAGAATGGTCCTGCACCAGTAATA
___________________________________________ 901 Left GACATGTTCTAATATAGTCACATTTT
4883 Left CATGTTCTAATATAGTCACATTTTCA 4 CATT
4884 Right AGAATGGTCCTGCACCAGTAATA 4902 Right GAATGGTCCTGCACCAGTAATATG
Seq. .
4 ACATGTTCTAATATAGTCACATTTTC ID Pnmer Sequence 885 Left A 901 4 Left GACATGTTCTAATATAGTCACATTTT
4886 Right AGAATGGTCCTGCACCAGTAATA CATT
4902 Right GAATGGTCCTGCACCAGTAATATG
GACATGTTCTAATATAGTCACATTTT
____________________________________________________________ 4887 Left CA 903 L eft ACATGTTCTAATATAGTCACATTTTC

4888 Right GAATGGTCCTGCACCAGTAATATG A
4904 Right GAATGGTCCTGCACCAGTAATATG
CATGTTCTAATATAGTCACATTTTCAT
4889 Left ___________________________________________ 905 4 Left GACATGTTCTAATATAGTCACATTTT
4890 Right AAGAATGGTCCTGCACCAGTAATA CAT
4906 Right GAATGGTCCTGCACCAGTAATATG
CATGTTCTAATATAGTCACATTTTCAT
4891 Left ___________________________________________ 907 L eft CATGTTCTAATATAGTCACATTTTCAT

4892 Right AGAATGGTCCTGCACCAGTAATA _______ 4908 Right GAATGGTCCTGCACCAGTAATATG
4893 Left CATGTTCTAATATAGTCACATTTTCAT
4894 Right AAGAATGGTCCTGCACCAGTAATA 4909 Left CATGTTCTAATATAGTCACATTTTCA
4910 Right AGAATGGTCCTGCACCAGTAAT
4895 Left CATGTTCTAATATAGTCACATTTTCAT
4896 Right AGAATGGTCCTGCACCAGTAATA 4911 Left TGACATGTTCTAATATAGTCACATTT
TCAT
4897 Left CATGTTCTAATATAGTCACATTTTCA 4912 Right ATGGTCCTGCACCAGTAATATG
4898 Right GAATGGTCCTGCACCAGTAATATG ______ ACATGTTCTAATATAGTCACATTTTC
__________________________________________ < 913 Left A

4914 Right AGAATGGTCCTGCACCAGTAAT TCAT
4918 Right ATGGTCCTGCACCAGTAATATGC
4915 Left CATGTTCTAATATAGTCACATTTTCAT
4916 Right GAATGGTCCTGCACCAGTAATATG
TGACATGTTCTAATATAGTCACATTT
4919 Left TCAT
4917 Left TGACATGTTCTAATATAGTCACATTT 4920 Right AATGGTCCTGCACCAGTAATATGC
KRAS Exon2 211-300 bases Seq. Primer Sequence ID 1941 Left TTAAAAGGTACTGGTGGAGTATTTGA
4921 Left AAAAGGTACTGGTGGAGTATTTGAT 1942 Right TACTCATGAAAATGGTCAGAGAAAC
4922 Right ATGAAAATGGTCAGAGAAACCTTTA _____ Left AGGTACTGGTGGAGTATTTGATAGTG
Seq. 4 943 Primer Se quence ID 1944 Right ATGAAAATGGTCAGAGAAACCTTTA
4923 Left TTAAAAGGTACTGGTGGAGTATTTGA
4924 Right ATGAAAATGGTCAGAGAAACCTTTA Left ATTAAAAGGTACTGGTGGAGTATTTG
'1945 A
4925 Left TAAAAGGTACTGGTGGAGTATTTGA 1946 Right ATGAAAATGGTCAGAGAAACCTTTA
4926 Right ATGAAAATGGTCAGAGAAACCTTTA
'1947 Left TTTGTATTAAAAGGTACTGGTGGAGT
4927 Left GGTACTGGTGGAGTATTTGATAGTG 1948 Right ATGAAAATGGTCAGAGAAACCTTTA
4928 Right ATGAAAATGGTCAGAGAAACCTTTA
4 949 Left GTTTGTATTAAAAGGTACTGGTGGAG
4929 Left AGGTACTGGTGGAGTATTTGATAGTG 4 950 Right ATGAAAATGGTCAGAGAAACCTTTA
4930 Right ATGAAAATGGTCAGAGAAACCTTTA
'1951 Left AAAAGGTACTGGTGGAGTATTTGATA
4931 Left TTTGTATTAAAAGGTACTGGTGGAG 4 952 Right ATGAAAATGGTCAGAGAAACCTTTA
4932 Right ATGAAAATGGTCAGAGAAACCTTTA
4 953 Left TAAAAGGTACTGGTGGAGTATTTGA
Left AAGGTACTGGTGGAGTATTTGATAGT '1954 Right TACTCATGAAAATGGTCAGAGAAAC

4934 Right ATGAAAATGGTCAGAGAAACCTTTA 4 955 Left TTGTATTAAAAGGTACTGGTGGAGT
'1956 Right ATGAAAATGGTCAGAGAAACCTTTA
4935 Left GGTACTGGTGGAGTATTTGATAGTGT
4936 Right ATGAAAATGGTCAGAGAAACCTTTA 4 957 Left GGTACTGGTGGAGTATTTGATAGTG
'1958 Right TACTCATGAAAATGGTCAGAGAAAC
4937 Left AAAAGGTACTGGTGGAGTATTTGAT
4938 Right TACTCATGAAAATGGTCAGAGAAAC '1959 Left AAAGGTACTGGTGGAGTATTTGATA
'1960 Right ATGAAAATGGTCAGAGAAACCTTTA
4939 Left GTTTGTATTAAAAGGTACTGGTGGA
4940 Right ATGAAAATGGTCAGAGAAACCTTTA
KRAS Exon2 301-400 bases Seq. Pn .
mer Sequence ID 4967 Left AAAAGGTACTGGTGGAGTATTTGAT
4961 Left AAAAGGTACTGGTGGAGTATTTGAT 4968 Right TAACTTGAAACCCAAGGTACATTTC
4962 Right CCAAGGAAAGTAAAGTTCCCATATT
4969 Left AAAAGGTACTGGTGGAGTATTTGAT
4963 Left TTAAAAGGTACTGGTGGAGTATTTGA 4970 Right GAAACCCAAGGTACATTTCAGATAA
4964 Right CCAAGGAAAGTAAAGTTCCCATATT
__________________________________________ 4971 Left TTAAAAGGTACTGGTGGAGTATTTGA
4965 Left TAAAAGGTACTGGTGGAGTATTTGA 4972 Right TAACTTGAAACCCAAGGTACATTTC
4966 Right CCAAGGAAAGTAAAGTTCCCATATT

4973 Left GGTACTGGTGGAGTATTTGATAGTG
4974 Right CCAAGGAAAGTAAAGTTCCCATATT 487 Left GGTACTGGTGGAGTATTTGATAGTGT
4988 Right CCAAGGAAAGTAAAGTTCCCATATT
Seq.
Primer Sequence ID
4975 Left TGAAGTACAGTTCATTACGATACACG 4989 Left GGTACTGGTGGAGTATTTGATAGTG
4976 Right ATGAAAATGGTCAGAGAAACCTTTA 4990 Right TAACTTGAAACCCAAGGTACATTTC
4977 Left TAAAAGGTACTGGTGGAGTATTTGA 4991 Left ATTACGATACACGTCTGCAGTCAAC
4978 Right TAACTTGAAACCCAAGGTACATTTC 4992 Right ATGAAAATGGTCAGAGAAACCTTTA
4979 Left AGGTACTGGTGGAGTATTTGATAGTG 4993 Left ACAGTTCATTACGATACACGTCTGC
4980 Right CCAAGGAAAGTAAAGTTCCCATATT 4994 Right ATGAAAATGGTCAGAGAAACCTTTA
4981 Left CAGTCAACTGGAATTTTCATGATT 4995 Left TACAGTTCATTACGATACACGTCTG
4982 Right ATGAAAATGGTCAGAGAAACCTTTA 4996 Right ATGAAAATGGTCAGAGAAACCTTTA
4983 Left TTAAAAGGTACTGGTGGAGTATTTG 4997 Left TGGAGGAGTTTGTAAATGAAGTACAG
4984 Right CCAAGGAAAGTAAAGTTCCCATATT 4998 Right TTTATCTGTATCAAAGAATGGTCCTG
4985 Left AAGGTACTGGTGGAGTATTTGATAGT 99 Left AGGTACTGGTGGAGTATTTGATAGTG
4986 Right CCAAGGAAAGTAAAGTTCCCATATT Sc, 00 Right TAACTTGAAACCCAAGGTACATTTC
KRAS Exon2 401-600 bases Seq. Pn .
mer Sequence ID 5019 Left AGTCATGATATGATCCTTTGAGAGC

5001 Left AGTCATGATATGATCCTTTGAGAGC 5020 Right TTTATCTGTATCAAAGAATGGTCCTG
5002 Right ATGAAAATGGTCAGAGAAACCTTTA
Seq. Primer Se quence 5003 Left ATATGATCCTTTGAGAGCCTTTAGC ID
5004 Right GAAACCCAAGGTACATTTCAGATAA 5021 Left AGTCATGATATGATCCTTTGAGAGC
5022 Right TACTCATGAAAATGGTCAGAGAAAC
5005 Left TGATATGATCCTTTGAGAGCCTTTA
5006 Right ATGAAAATGGTCAGAGAAACCTTTA 5023 Left TGGAGGAGTTTGTAAATGAAGTACAG
5024 Right TAACTTGAAACCCAAGGTACATTTC
5007 Left ATATGATCCTTTGAGAGCCTTTAGC
5008 Right ATGAAAATGGTCAGAGAAACCTTTA 5025 Left TGGAGGAGTTTGTAAATGAAGTACAG
5026 Right GAAACCCAAGGTACATTTCAGATAA
5009 Left TGAAGTCATGATATGATCCTTTGAG
5010 Right ATGAAAATGGTCAGAGAAACCTTTA 5027 Left AGTCATGATATGATCCTTTGAGAGC
5028 Right TTATCTGTATCAAAGAATGGTCCTG
5011 Left TGGAGGAGTTTGTAAATGAAGTACAG
5012 Right CCAAGGAAAGTAAAGTTCCCATATT 5029 Left ATATGATCCTTTGAGAGCCTTTAGC
5030 Right TGAAACCCAAGGTACATTTCAGATA
5013 Left TGGAGGAGTTTGTAAATGAAGTACAG
5014 Right TGACATACTCCCAAGGAAAGTAAAG 5031 Left GATATGATCCTTTGAGAGCCTTTAG
5032 Right GAAACCCAAGGTACATTTCAGATAA
5015 Left TGGAGGAGTTTGTAAATGAAGTACAG
5016 Right CTGACATACTCCCAAGGAAAGTAAA 5033 Left TGAAGTACAGTTCATTACGATACACG
5034 Right CCAAGGAAAGTAAAGTTCCCATATT
5017 Left ATGATATGATCCTTTGAGAGCCTTT
5018 Right ATGAAAATGGTCAGAGAAACCTTTA 5035 Left TGAAGTACAGTTCATTACGATACACG

5036 Right CGAAACTCTGAAATACACTTCCAAT
5039 Left TGAAGTACAGTTCATTACGATACACG
5037 Left TGAAGTACAGTTCATTACGATACACG 5040 Right TGACATACTCCCAAGGAAAGTAAAG
5038 Right CTGACATACTCCCAAGGAAAGTAAA
KRAS Exon2 601-800 bases Seq. . 5059 Left AGTCATGATATGATCCTTTGAGAGC
Pnmer Sequence ID 5060 Right GAAACCCAAGGTACATTTCAGATAA
5041 Left AGTCATGATATGATCCTTTGAGAGC
5042 Right CCAAGGAAAGTAAAGTTCCCATATT 5061 Left CTACTGCCATGATGCTTTAAAAGTT
5062 Right TAACTTGAAACCCAAGGTACATTTC
5043 Left AGTCATGATATGATCCTTTGAGAGC
5044 Right CGAAACTCTGAAATACACTTCCAAT 5063 Left AGAGCACTGTGAAGTCTCTACATGA
5064 Right CCAAGGAAAGTAAAGTTCCCATATT
Seq. . 5065 Left AGTCATGATATGATCCTTTGAGAGC
Pnmer Sequence ID 5066 Right TACAAATTTCTACCCTCTCACGAAA
5045 Left AGTCATGATATGATCCTTTGAGAGC
5046 Right TGACATACTCCCAAGGAAAGTAAAG 5067 Left AGTCATGATATGATCCTTTGAGAGC
5068 Right AAATTTCTACCCTCTCACGAAACTC
5047 Left AGTCATGATATGATCCTTTGAGAGC
5048 Right CTGACATACTCCCAAGGAAAGTAAA 5069 Left CTACTGCCATGATGCTTTAAAAGTT
5070 Right GAAACCCAAGGTACATTTCAGATAA
__________________________________________ 5071 Left AGAGCACTGTGAAGTCTCTACATGA
5049 Left AGTCATGATATGATCCTTTGAGAGC 5072 Right CTGACATACTCCCAAGGAAAGTAAA
5050 Right AATTTCTACCCTCTCACGAAACTCT
__________________________________________ 5073 Left AGAGCACTGTGAAGTCTCTACATGA
5051 Left AGTCATGATATGATCCTTTGAGAGC 5074 Right TGACATACTCCCAAGGAAAGTAAAG
5052 Right GATACAAATTTCTACCCTCTCACGA ____ 5075 Left TCTGTAGCTGTTGCATATTGACTTC
5053 Left TCTGTAGCTGTTGCATATTGACTTC 5076 Right TAACTTGAAACCCAAGGTACATTTC
5054 Right CCAAGGAAAGTAAAGTTCCCATATT ____ 5077 Left AGAGCACTGTGAAGTCTCTACATGA
5055 Left TTTTTCTGTAGCTGTTGCATATTGA 5078 Right AATTTCTACCCTCTCACGAAACTCT
5056 Right CCAAGGAAAGTAAAGTTCCCATATT ____ D79 Left AGTCATGATATGATCCTTTGAGAGC
5057 Left AGTCATGATATGATCCTTTGAGAGC 5 D80 Right ACGAAACTCTGAAATACACTTCCAA
5058 Right TAACTTGAAACCCAAGGTACATTTC
KRAS Exon2 801-1000 bases Seq. . 5088 Right TGACATACTCCCAAGGAAAGTAAAG
ID Pnmer Sequence 5081 Left ATCCAGCTTTATTTGACACTCATTC 5089 Left AATATTGTTCTTCTTTGCCTCAGTG
5082 Right CCAAGGAAAGTAAAGTTCCCATATT 5090 Right CTGACATACTCCCAAGGAAAGTAAA
5083 Left CTACTGCCATGATGCTTTAAAAGTT 5091 Left ATCCAGCTTTATTTGACACTCATTC
5084 Right CCAAGGAAAGTAAAGTTCCCATATT 5092 Right TGACATACTCCCAAGGAAAGTAAAG
5085 Left ATCCAGCTTTATTTGACACTCATTC 5093 Left ATCCAGCTTTATTTGACACTCATTC
5086 Right CGAAACTCTGAAATACACTTCCAAT 5094 Right CTGACATACTCCCAAGGAAAGTAAA
5087 Left AATATTGTTCTTCTTTGCCTCAGTG 5095 Left CTACTGCCATGATGCTTTAAAAGTT

5096 Right CGAAACTCTGAAATACACTTCCAAT 5108 Right CGAAACTCTGAAATACACTTCCAAT
5097 Left CTACTGCCATGATGCTTTAAAAGTT 5109 Left TCTGTAGCTGTTGCATATTGACTTC
5098 Right TGACATACTCCCAAGGAAAGTAAAG 510 Right TGACATACTCCCAAGGAAAGTAAAG
Seq.
Primer Sequence ID 5111 Left TCTGTAGCTGTTGCATATTGACTTC

5099 Left CTACTGCCATGATGCTTTAAAAGTT 5112 Right AATTTCTACCCTCTCACGAAACTCT
5100 Right CTGACATACTCCCAAGGAAAGTAAA
5113 Left TCTGTAGCTGTTGCATATTGACTTC
5101 Left ATCCAGCTTTATTTGACACTCATTC 5114 Right GATACAAATTTCTACCCTCTCACGA
5102 Right AATTTCTACCCTCTCACGAAACTCT
5115 Left AAAAGTTTTTCTGTAGCTGTTGCAT
5103 Left ATCCAGCTTTATTTGACACTCATTC 5116 Right CCAAGGAAAGTAAAGTTCCCATATT
5104 Right GATACAAATTTCTACCCTCTCACGA
5117 Left TTTTTCTGTAGCTGTTGCATATTGA
5105 Left CTACTGCCATGATGCTTTAAAAGTT 5118 Right CGAAACTCTGAAATACACTTCCAAT
5106 Right GATACAAATTTCTACCCTCTCACGA
5119 Left TTTTTCTGTAGCTGTTGCATATTGA
5107 Left TCTGTAGCTGTTGCATATTGACTTC 5120 Right TGACATACTCCCAAGGAAAGTAAAG
KRAS Exon2 2kb Primer Sequence Primer Sequence ID ID
5121 Left AGTCATGATATGATCCTTTGAGAGC 5141 Left TCTTCAGTCAATTATGATGCTGTGT
5122 Right TCCTAACCCACTTTATCACATTCAT 5142 Right CCAAGGAAAGTAAAGTTCCCATATT
5123 Left AGTCATGATATGATCCTTTGAGAGC 5143 Left GTCTTCAGTCAATTATGATGCTGTG
5124 Right AATTAGACTGTTCCCCTTTACTGCT 5144 Right CCAAGGAAAGTAAAGTTCCCATATT
5125 Left ATTATGTGTTACATGAGGGAAAGGA 5145 Left AATATTGTTCTTCTTTGCCTCAGTG
5126 Right CCAAGGAAAGTAAAGTTCCCATATT 5146 Right AATTAGACTGTTCCCCTTTACTGCT
5127 Left AGTCATGATATGATCCTTTGAGAGC 5147 Left ATCCAGCTTTATTTGACACTCATTC
5128 Right TTGGAAACAAAGTGTAATGGAATTT 5148 Right AATTAGACTGTTCCCCTTTACTGCT
5129 Left AGTCATGATATGATCCTTTGAGAGC 5149 Left CTACTGCCATGATGCTTTAAAAGTT
5130 Right TCAATTAGACTGTTCCCCTTTACTG 5150 Right TCTTTGCAAATAGGCATTATTTCTC
5131 Left ATTATGTGTTACATGAGGGAAAGGA 5151 Left ATTATGTGTTACATGAGGGAAAGGA
5132 Right CGAAACTCTGAAATACACTTCCAAT 5152 Right GATACAAATTTCTACCCTCTCACGA
5133 Left ATTATGTGTTACATGAGGGAAAGGA 5153 Left GCACATTCATTAATTTGGAGCTACT
5134 Right TGACATACTCCCAAGGAAAGTAAAG 5154 Right CTGACATACTCCCAAGGAAAGTAAA
5135 Left ATTATGTGTTACATGAGGGAAAGGA 5155 Left CTACTGCCATGATGCTTTAAAAGTT
5136 Right CTGACATACTCCCAAGGAAAGTAAA 5156 Right AATTAGACTGTTCCCCTTTACTGCT
5137 Left AATATTGTTCTTCTTTGCCTCAGTG 5157 Left AATATTGTTCTTCTTTGCCTCAGTG
5138 Right TCTTTGCAAATAGGCATTATTTCTC 5158 Right ACCAAAAATATGTGACGTTTCCTTA
5139 Left ATCCAGCTTTATTTGACACTCATTC 5159 Left AATATTGTTCTTCTTTGCCTCAGTG
5140 Right TCTTTGCAAATAGGCATTATTTCTC 5160 Right TACCAAAAATATGTGACGTTTCCTT

KRAS Exon2 5kb S eq. .
Pnmer Sequence ID 5181 Left TCCTCATCTATTCCTAATTGCTTTG

5161 Left CAGCCAATAAGTCTAGGTAGAGCAG M82 Right TTGAACTGAATTATAAGTGCCACAA
5162 Right TAAAGATGAAACAAACCAATCCAAT
M83 Left TTTGCTTTTAAGAGATGGTAGATGG
5163 Left AGCCTTCTTAAATCTTGAGATCCAT 5184 Right TCCTAACCCACTTTATCACATTCAT
5164 Right TCTTTGCAAATAGGCATTATTTCTC ___ 5185 Left GCACATTCATTAATTTGGAGCTACT
5165 Left ATTATGTGTTACATGAGGGAAAGGA M86 Right TAAAGATGAAACAAACCAATCCAAT
5166 Right TAAAGATGAAACAAACCAATCCAAT
5187 Left GACTTAAACATGTGCATCTCCTTTT
5167 Left GGCTAGTAAACTTTTTGGCCTTAAC M88 Right AAATGACAACAAAGCAAAGGTAAAG
5168 Right TCTTTGCAAATAGGCATTATTTCTC
5189 Left TCCTCATCTATTCCTAATTGCTTTG
5169 Left AGCCTTCTTAAATCTTGAGATCCAT 5190 Right CCTTACTGAATAGGAAACTGTTC CA
5170 Right AATTAGACTGTTCCCCTTTACTGCT
S eq. . M91 Left TTTGCTTTTAAGAGATGGTAGATGG
Pnmer Sequence ID M92 Right AATTAGACTGTTCCCCTTTACTGCT

5171 Left GGCTAGTAAACTTTTTGGCCTTAAC _____ 5172 Right TCCTAACCCACTTTATCACATTCAT 5193 Left AGTCATGATATGATCCTTTGAGAGC
5194 Right AATTAGCATGATTGCCTAGAAACAC
5173 Left GGCTAGTAAACTTTTTGGCCTTAAC _____ 5174 Right AATTAGACTGTTCCCCTTTACTGCT 5195 Left AGCCTTCTTAAATCTTGAGATCCAT
5196 Right ACCAAAAATATGTGACGTTTCCTTA
5175 Left TTTGCTTTTAAGAGATGGTAGATGG _____ 5176 Right TCTTTGCAAATAGGCATTATTTCTC 5197 Left AGCCTTCTTAAATCTTGAGATCCAT
5198 Right TACCAAAAATATGTGACGTTTCCTT
5177 Left CAGCCAATAAGTCTAGGTAGAGCAG ______ 5178 Right TTGAACTGAATTATAAGTGCCACAA 5199 Left AGCCTTCTTAAATCTTGAGATCCAT
5200 Right TCAATTAGACTGTTCCCCTTTACTG
5179 Left GGCTAGTAAACTTTTTGGCCTTAAC
5180 Right TTGGAAACAAAGTGTAATGGAATTT
Table 11. ALK cDNA Capture Primer List for NGS Panel ALK Regionl 75-125 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
5201 Left ctgcacactggccgtct 5205 Left actgcacactgg 5202 Right aaccatgatccctggagtg 5206 Right accatgatccct 5203 Left actgcacactggccg 5204 Right accatgatccctgga ALK Regionl 126-175 bases Seq. . 5209 Left gactccaagcacaccatcct Primer Sequence ID 5210 Right aaccatgatccctggagtg 5207 Left acaccatcctgagtccgtg 5208 Right cactgtccaaccatgatcc 5211 Left cctgagtccgtggatgagg f, 212 Right ctggagcactgtccaac cat 5221 Left atgaggagcagcagtgag 5213 Left tccaagcacaccatcctgag 5222 Right gattcttccctggagcactgtccaa 5214 Right ccaaccatgatccctgga 5223 Left catcctgagtccgtggat 5215 Left ctgcacactggccgtct 5224 Right agcactgtccaaccatgct 5216 Right ctcgaaatgggttgtctggac 5225 Left tgagcactgcacact 5217 Left ccgtggatgaggagcagc 5226 Right cc cgattcttccctggagc 5218 Right gattcttccctggagcactgtc Seq. . 5227 Left atgaggagcagcagt Primer Sequence ID 5228 Right gacgcccgattcttccct 5219 Left gagcactgcacactggc 5220 Right cgattcttccctggagcact 229 Left gtgagcactgcac __________________________________________ 4230 Right aatgggttgtctggacgcc ALK Regionl 76-225 bases Seq.
Primer Sequence ID 5251 Left accatcctgagtccgtggat 5231 Left ctectnctecttctcaacacct 5252 Right ggccactcgaaatgggttg 5232 Right attcttccctggagcactgtc Seq. . 5253 Left ccgtggatgaggagcagc Pnmer Sequence ID 5254 Right ggagatgtattccagggcca 5233 Left cttctcaacacctcagctgact 5234 Right cgattcttccctggagcact 5255 Left gagcactgcacactggc 5235 Left tactccttctcaacacctcagct 5256 Right gacaagctuggtttccac 5236 Right gcactgtccaaccatgcttc 5257 Left gcacaccatcctgagtcc 5237 Left ctgactccaagcacaccatc 5258 Right gaaatgggttgtctggacgcc 5238 Right ctcgaaatgggttgtctggac 5259 Left atgaggagcagcagtgag 5239 Left tccaagcacaccatcctgag 5260 Right agggccactcgaaatggg 5240 Right ccactcgaaatgggttgtctg 5261 Left ccgtggatgaggagc 5241 Left cctgagtccgtggatgagg 5262 Right cc cgattcttccctggagc 5242 Right gagatgtattccagggccactc 5263 Left tgagcactgcacact 5243 Left ttctcaacacctcagctgactccaa 5264 Right aagctuggtttccactgg 5244 Right gattcttccctggagcactgtccaa 5265 Left atgaggagcagcagt 5245 Left gaggctectnctecttctca 5266 Right ggacgcccgattcttccc 5246 Right ctggagcactgtccaacca 5267 Left gtgagcactgcac 5247 Left ctcagctgactccaagcaca 5268 Right tccactggagatgtattcca 5248 Right gagcactgtccaaccatgc 5269 Left ttctccttctcaaca 5249 Left acaccatcctgagtccgtg 5270 Right gcccgattcttccctgg 5250 Right ttccagggccactcgaaat ALK Regionl 226-275 bases Seq. 5273 Left gaggctectttctccttctcaa Primer Sequence ID 5274 Rjght ccactcgaaatgggttgtctg 5271 Left ctectnctecttctcaacacct 5272 Right ctcgaaatgggttgtctggac 5275 Left cttctcaacacctcagctgactc ___________________________________________ 276 Right gagatgtattccagggccactc 5293 Left ctcagctgactccaagcaca 5277 Left acaccatcctgagtccgtg 5294 Right gaaatgggttgtctggacgcc 5278 Right caaagaagtccactgcagacaag 5295 Left ccgtggatgaggagcagc 5279 Left ctcatctcaacacctcagct 5296 Right ctgcagttatcagggcaaag 5280 Right ggagatgtattccagggcca 5297 Left accatcctgagtccgtggat 5281 Left c cagaggctc Mete cttc 5298 Right ggcaaagaagtccactgca 5282 Right ttccagggccactcgaaat 5299 Left atgaggagcagcagtgag 5283 Left cctgagtccgtggatgagg 5300 Right chcactgcagttatcagggc 5284 Right gcaaagaagtccactgcagac 5301 Left cc cagaggctectnctcc 5285 Left ttctcaacacctcagctgactccaa 5302 Right gggccactcgaaatggg 5286 Right ggccactcgaaatgggttg Seq. . 5303 Left gcacaccatcctgagtcc Primer Sequence ID 5311 Rjght tccactggagatgtattcca 5287 Left gagcactgcacactggc 5288 Right gttcatcactgcagttcttcag 5305 Left gtgagcactgcaca 5306 Ri ht at ttcatcactgcagttat 5289 Left ctgactccaagcacaccatc 5290 Right caagctuggtttccactg 5307 Left ccgtggatgaggagc 5308 Right ttcagggcaaagaagtcc 5291 Left tccaagcacaccatcctgag 5292 Right agacaagctuggthcca 5309 Left tgaggagcagcagt __________________________________________ 5310 Right tgacchcactgcagtt ALK Region2 75-125 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
5311 Left cactccagggaagcatggt 5323 Left cactccagggaagcat 5312 Right caaagaagtccactgcagacaag 5324 Right caaagaagtccactgcagac 5313 Left aacgaggctgcaagagagat 5325 Left tectectgatgccca 5314 Right gagatgtattccagggccact 5326 Right agacaagctuggttt 5315 Left gaggctgcaagagagatcct 5327 Left ccactccagggaag 5316 Right tggagatgtattccagggcc 5328 Right aaagaagtccactgca 5317 Left ctcctgatgcccactccag 5329 Left acgaggctgcaag 5318 Right caagctgeggtttccactg 5330 Right tccactggagatgtatt 5319 Left tectectgatgcccactc 5331 Left agagatc ctc ctga 5320 Right agacaagctuggtttcca 5332 Right tccactggagatgt 5321 Left acgaggctgcaagaga 5322 Right tccactggagatgtattcca ALK Region2 126-175 bases Seq.
Primer Sequence ID .5335 Left ctcctgatgcccactccag 5333 Left aacgaggctgcaagagagat 5336 Ri ht at ttecttcactgcagttat 5334 Right caaagaagtccactgcagacaag 5337 Left gaggctgcaagagagatcct 5352 Right agacaagctuggttt 5338 Right cttcactgcagttcttcaggg 5353 Left tectectgatgccca 5339 Left cacaacgaggctgcaagag 5354 Right gatgaccttcactgcagtt 5340 Right gcaaagaagtccactgcagac 5355 Left gccccacaacgaggct 5341 Left ctctggaaggtacattgcccag 5356 Right tccactggagatgtattc 5342 Right caagctgeggtttccactg 5357 Left gctgctgccccacaac 5343 Left gaaggtacattgcccagctg 5358 Right ggcaaagaagtccact 5344 Right agacaagctuggtttcca 5359 Left ctctggaaggtacattgcc 5345 Left tectectgatgcccactc 5360 Right agacaagctugg 5346 Right ctgcagttcttcagggcaaag 5361 Left cacctgcagccctct 5347 Left cccacaacgaggctgcaa 5362 Right tccactggagatgta 5348 Right ggcaaagaagtccactgca Seq. 5363 Left agagatc ctc ctga Primer Sequence ID .5364 Ri ht at ttccttcactgca 5349 Left tgctgccccacaacgag 5365 Left ctgctgccccac 5350 Right ttcagggcaaagaagtcc 5366 Rjght tcagggcaaagaag 5351 Left gtacattgcccagctgctg ALK Region2 176-225 bases Seq. 5385 Left gagcactgcacactggc Primer Sequence ID 5386 Right gacaagctuggatccac 5367 Left gaaggtacattgcccagctg 5368 Right gatgttc cttcactgcagttctt Seq.
ID Primer Sequence 5369 Left gaggctgcaagagagatcct 5387 Left actgcacactggccgtc 5370 Right gttccttcactgcagttcttcag 5388 Right aagctuggtaccactgg 5371 Left gtacattgcccagctgctg 5389 Left tgctgccccacaacgag 5372 Right caaagaagtccactgcagacaag 5390 Right gctctgcagggccatct 5373 Left ctcctgatgcccactccag 5391 Left gccccacaacgaggct 5374 Right cattccaacaagtgaaggagctc 5392 Right gatgaccttcactgcagtt 5375 Left ctctggaaggtacattgccca 5393 Left cacctgcagccctct 5376 Right actgcagttcttcagggcaa 5394 Right gcaaagaagtccactgcagac 5377 Left cacaacgaggctgcaagagagat 5395 Left tectectgatgccca 5378 Right ctgcagttcttcagggcaaaga 5396 Right gactgtc ccattc caacaagtg 5379 Left tectectgatgcccactc 5397 Left ctctggaaggtacattgc 5380 Right cc cattccaacaagtgaaggag 5398 Right ggcaaagaagtccactgca 5381 Left acaacgaggctgcaagaga 5399 Left gctgctgccccacaac 5382 Right atcttggagc ctggggatgttc 5400 Right ttcagggcaaagaagtcc 5383 Left ccacaacgaggctgcaag 5401 Left cgtctcggtgcacagg 5384 Right atcttggagcctggggatg 5402 Right agacaagctuggtttc 5403 Left ccgtggatgaggagcagc 5405 Left agagatc ctc ctga 5404 Right tccactggagatgtattc 5406 Right caacaagtgaaggagctctgc ALK Region2 226-275 bases Seq. 5426 Right agacaagctuggthcca ID Primer Sequence 5407 Left ctctggaaggtacattgcccag 5427 Left ccgtggatgaggagcagc 5408 Right cattccaacaagtgaaggagctc 5428 Right ctgcagttatcagggcaaag 5409 Left aacgaggctgcaagagagatc 5429 Left accatcctgagtccgtggat 5410 Right cc cattccaacaagtgaaggag 5430 Right ggcaaagaagtccactgca 5411 Left acaccatcctgagtccgtg 5431 Left tcct . at .
cccactcca 5412 Right caaagaagtccactgcagacaag 5432 Rjght ctcatcttctccctgggca 5413 Left gaaggtacattgcccagctg 5433 Left cacaacgaggctgcaagag 5414 Right caacaagtgaaggagctctgc 5434 Right ggaagtcacaggcctgcc 5415 Left gtacattgcccagctgctg 5435 Left atgaggagcagcagtgag 5416 Right gactgtcc cattccaacaagtg 5436 Right cttcactgcagttatcagggc Seq. 5437 Left ctctggaaggtacattgcc Primer Sequence ID .5438 Right atchggagcctggggatgttc 5417 Left ctgcacactggccgtct 5418 Right gatgttc chcactgcagttat 5439 Left ctgctgccccacaacga ___________________________________________ 5440 Right tgaaggagctctgcaggg 5419 Left cctgagtccgtggatgagg 5420 Right gcaaagaagtccactgcagac 5441 Left cgtctcggtgcacagg ___________________________________________ 5442 Right atcttggagcctggggatg 5421 Left gagcactgcacactggc 5422 Right gttcatcactgcagttatcag 5443 Left gccccacaacgaggct ___________________________________________ 5444 Right gaaggagctctgcagggccatc 5423 Left ctgactccaagcacaccatc 5424 Right caagctgeggtttccactg 5445 Left cccacaacgaggctgcaa ___________________________________________ 5446 Right gactgtcc cattc caacaa 5425 Left tccaagcacaccatcctgag ALK Region3 76-125 bases Seq. 5456 Right gctctgcagggccatct Primer Sequence ID Seq.
Primer Sequence 5447 Left ggaatacatctccagtggaaacc ID
5448 Right cattccaacaagtgaaggagctc 5457 Left catttcgagtggccctgga 5458 Right gagctctgcagggcca 5449 Left gtccagacaacccatttcgag 5450 Right atcttggagcctggggatg 5459 Left gcgtccagacaacccatttc 5460 Right atcttggagcctgggg 5451 Left tggccctggaatacatctcc 5452 Right acaagtgaaggagctctgca 5461 Left tggaatacatctccagtggaa 5462 Right attccaacaagtgaaggag 5453 Left cgagtggccctggaatacatc 5454 Right tgaaggagctctgcaggg 5463 Left ggcgtccagacaacccat 5464 Right catcttggagcctg 5455 Left tcgagtggccctggaatac 5465 Left ggaatacatctccagtg 5466 Right cattccaacaagtgaag ALK Region3 126-175 bases Seq. 5481 Left gaagaatcgggcgtccaga Primer Sequence ID .5482 Right gactgtcc cattc caacaa 5467 Left gtccagacaacccatttcgag 5468 Right cattccaacaagtgaaggagctc 5483 Left tcgagtggccctggaata 5484 Right ggaagtcacaggcctgcc 5469 Left gagtggccctggaatacatctc 5470 Right cc cattccaacaagtgaaggag 5485 Left catttcgagtggccctgga __________________________________________ 5486 Right cccattccaacaagtgaag 5471 Left cgagtggccctggaatacat 5472 Right gactgtcc cattccaacaagtg 5487 Left ggcgtccagacaacccat 5488 ht caagctggaggactgtc 5473 Left gacagtgctccagggaagaat 5474 Right caacaagtgaaggagctctgc 5489 Left gaagaatcgggcgtccagacaa 5490 Right gactgtcc cattc caa 5475 Left ggaagcatggttggacagtg _5491 Left ccatttcgagtggccct 5476 Right gctctgcagggccatct 5492 Rjght caagctggaggact 5477 Left tgctccagggaagaatcgg 5493 Left ggaagaatcgggcgtcc 5478 Right tgaaggagctctgcaggg 5494 Right ggactgtcc cattc Seq. 5495 Left ccatttcgagtggc Primer Sequence ID .5496 Right ggcctgcccaag 5479 Left gcgtccagacaacccatttc 5480 Right gagctctgcagggcca ALK Region3 76-225 bases Seq. 5512 Right ctctcatcttctccctgggc ID Primer Sequence 5497 Left gagtggccctggaatacatctc 5513 Left aacgaggctgcaagagagat 5498 Right acatctggctctcatcttctcc 5514 Right gctctgcagggccatct 5499 Left gacagtgctccagggaagaat 5515 Left gcatggaggacagtgctc 5500 Right cattccaacaagtgaaggagctc 5516 Right tgaaggagctctgcaggg 5501 Left ggaagcatggttggacagtg Seq.
Primer Sequence 5502 Right cc cattccaacaagtgaaggag ID
5517 Left atggaggacagtgctccag 5503 Left ccagacaacccatttcgagtg 5518 Right ggaagtcacaggcctgcc 5504 Right atctggctctcatcttctccctg 5519 Left agggaagcatggttggaca 5505 Left cactccagggaagcatggtt 5520 Right gagctctgcagggcca 5506 Right gactgtcc cattccaacaagtg 5521 Left tgctccagggaagaatcgg 5507 Left gaggctgcaagagagatcct 5522 Right gactgtcc cattc caacaa 5508 Right caacaagtgaaggagctctgc 5523 Left tcgagtggccctggaat 5509 Left cgagtggccctggaataca 5524 Right gcagtttc cggcacatctg 5510 Right gcacatctggctctcatcttc 5525 Left ctcctgatgcccactccag 5511 Left cgtccagacaacccatttcg 5526 Right cccattccaacaagtgaag 5532 Right ttctccctgggcaca 5527 Left ccatttcgagtggccctgg 5528 Right gcacatctggctctcatc 5533 Left ggacagtgctccagggaag 5534 Right tggaagtcacaggcc 5529 Left gaagaatcgggcgtccaga 5530 Right caagctggaggactgtc 5535 Left agggaagaatcgggcgtc 5536 Right gactgtcc cattc caa 5531 Left gaagaatcgggcgtccagacaac ALK Region3 226-275 bases Seq.
Primer Sequence ID .5557 Left cgagtggccctggaatacat 5537 Left gagtggccctggaatacatctc 5558 Right tccagccacagaagccatc 5538 Right agccatchcaaagttgcagtaaaa 5559 Left atggaggacagtgctccag 5539 Left gtccagacaacccatttcgag 5560 Right gcagtttc cggcacatctg ¨
5540 Right aagccatchcaaagttgcagta _________________________________________ _5561 Left cacaacgaggctgcaagag 5541 Left ctctggaaggtacattgcccag 5562 Right ggaagtcacaggcctgcc 5542 Right cattccaacaagtgaaggagctc 5563 Left catttcgagtggccctgga 5543 Left gacagtgctccagggaagaatc 5564 Ri ht aa ccatchcaaagttgca 5544 Right acatctggctctcatcttctcc _ __________________________________________ Seq. 5565 Left gacagtgctccagggaaga Primer Sequence ID .5566 Right cacaggcagthccggc 5545 Left aacgaggctgcaagagagatc 5546 Right cc cattccaacaagtgaaggag 5567 Left cccacaacgaggctgcaa __________________________________________ 5568 Right tgaaggagctctgcaggg 5547 Left ggaagcatggttggacagtg 5548 Right gcacatctggctctcatcttc 5569 Left gcatggaggacagtgctc __________________________________________ 5570 Right gcacatctggctctcatc 5549 Left gaaggtacattgcccagctg 5550 Right caacaagtgaaggagctctgc 5571 Left tgctgccccacaacgag __________________________________________ 5572 Right agctctgcagggccatc 5551 Left gtacattgcccagctgctg 5552 Right gactgtcc cattccaacaagtg 5573 Left tcgagtggccctggaata __________________________________________ 5574 Right gtccagccacagaagcc 5553 Left gcgtccagacaacccatttc 5554 Right atctggctctcatcttctccctg 5575 Left gccccacaacgaggct __________________________________________ 5576 Right gactgtcc cattc caacaa 5555 Left cactccagggaagcatggtt 5556 Right ctctcatcttctccctgggc ALK Region4 226-275 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
5577 Left gagcactgcacactggc 5583 Left cgtctcggtgcac 5578 Right ttggagcctggggatgttc 5584 Right agctctgcagggcca 5579 Left ctgcacactggccgtct 5585 Left tgagcactgcacact 5580 Right atcttggagcctggggatg 5586 Right tggagcctgggg 5581 Left cgtctcggtgcacagg 5587 Left aggagcagcagtgag 5582 Right gctctgcagggccatct 5588 Right gatgttc cttcact ALK Region4 276-325 bases Seq. 5603 Left tgagcactgcacactgg Primer Sequence ID 5604 Ri ht ct cccattccaacaagtg 5589 Left ctgcacactggccgtct 5590 Right cattccaacaagtgaaggagctc 5605 Left cgtctcggtgcacagg 5606 Right tgaaggagctctgcaggg 5591 Left ctgactccaagcacaccatc 5592 Right atcttggagcctggggatgtt 5607 Left acaccatcctgagtccgtg 5608 Right atcttggagcctggg 5593 Left cactgcacactggccg 5594 Right cccattccaacaagtgaaggag 5609 Left tgagcactgcacac ______________________________________________ _5610 Right gactgtcc cattccaacaa 5595 Left atgaggagcagcagtgag 5596 Right caacaagtgaaggagctctgc 5611 Left cgtctcggtgcac 5612 ht cccattccaacaagtgaag 5597 Left cctgagtccgtggatgagg 5598 Right gctctgcagggccatct 5613 Left ttctcaacacctcagctgactc 5614 Right gatgttc cttcact 5599 Left tccaagcacaccatcctgag 5600 Right atcttggagcctggggat 5615 Left ccgtggatgaggagcagc 561Ri ht catcttggagcct 5601 Left ccatcctgagtccgtggat 5617 Left gtgagcactgca 5602 Right gagctctgcagggcca 5618 Right gactgtcc cattccaa Seq.
Primer Sequence ID
ALK Region4 326-375 bases Seq.
Primer Sequence ID 5635 Left cactgcacactggccgt 5619 Left cttctcaacac ctcagctgactc 636 Rjght ctctcatcttctccctgggc 5620 Right cattccaacaagtgaaggagctc 5637 Left cctgagtccgtggatgagg 5621 Left aacacctcagctgactccaag 5638 Right ggaagtcacaggcctgcc 5622 Right cc cattccaacaagtgaaggag ___________________________________________ Seq.
Primer Sequence 5623 Left tnctcatctcaacacctcagct ID
5624 Right caacaagtgaaggagctctgc 5639 Left gactccaagcacaccatcct 5640 Right gactgtcc cattc caacaa 5625 Left ctcagctgactccaagcaca 5626 Right gactgtcc cattccaacaagtg 5641 Left gaggctectnctecttctcaa 5642 Right atcttggagcctgggg 5627 Left cctnctecttctcaacacctca 5628 Right tatggagcctggggatg 5643 Left accatcctgagtccgtggat 5644 Right cccattccaacaagtgaag 5629 Left gctectnctccactcaacac 5630 Right agctctgcagggccatc 5645 Left tccaagcacaccatcctgag 5646 Right caagctggaggactgtc 5631 Left tgcacactggccgtctc 5632 Right ctggctctcatcttctccctg 5647 Left gagcactgcacactggc 5648 Right ttctccctgggcaca 5633 Left acaccatcctgagtccgtg 5634 Right tgaaggagctctgcaggg 5649 Left c cagaggctc Mete cttc 5650 Right gcagggccatcttg 5655 Left cccagaggctcattctcc 5651 Left gctgactc caagcacac cat 5656 Right catcttggagcctg 5652 Right gactgtc ccattccaa 5657 Left gcacaccatcctgagtcc 5653 Left ccgtggatgaggagcagc 5658 Right caagctggaggact 5654 Right tggaagtcacaggcc ALK Region4 376-425 bases Seq.
Primer Sequence ID 5679 Left cc cagaggctectnctcc 5659 Left ctectnctccactcaacacct 5680 Right aaggagctctgcagggc 5660 Right gactgtc ccattc caacaagtg 5681 Left ttctcaacacctcagctgactccaa 5661 Left gaggctectnctcatctcaac 5682 Right gactgtc ccattccaacaa 5662 Right cattccaacaagtgaaggagctc 5683 Left ccgtggatgaggagcagc 5663 Left ctcatctcaacacctcagct 5684 Right cacaggcagtttccggc 5664 Right cccattccaacaagtgaaggag 5685 Left ctcagctgactccaagcaca 5665 Left ccagaggctcattctecttc 5686 Rjght caagctggaggactgtc 5666 Right caacaagtgaaggagctctgc 5687 Left tggggcagagcgttct 5667 Left acaccatcctgagtccgtg 5688 Rjght cc cattccaacaagtgaag 5668 Right acatctggctctcatcttctcc Seq. . 5689 Left gcacaccatcctgagtcc Primer Sequence ID 5690 Right ttctccctgggcaca 5669 Left ctccaagcacaccatcctga 5670 Right ctggctctcatcttctccctg 5691 Left ccgtggatgaggagc __________________________________________ 5692 Right gcacatctggctctcatc 5671 Left accatcctgagtccgtggat 5672 Right gcacatctggctctcatcttc 5693 Left agagcgttctaaggagatg __________________________________________ 5694 Right gactgtc ccattccaa 5673 Left ctgactccaagcacaccatc 5674 Right ctctcatcttctccctgggc 5695 Left atgaggagcagcagt __________________________________________ 5696 Right gcagtttccggcacat 5675 Left cactcaacacctcagctgactc 5676 Right ggaagtcacaggcctgcc 5697 Left cc cagaggctectttc __________________________________________ 5698 Right caagctggaggact 5677 Left cctgagtccgtggatgagg 5678 Right gcagtttc cggcacatctg ALK Region4 426-475 bases Seq. b 705 Left cttctcaacacctcagctgactc Primer Sequence ID b 706 Right gcacatctggctctcatcttc 5699 Left ctectnctccactcaacacctc 5700 Right acatctggctctcatcttctcc b 707 Left acaccatcctgagtccgtg b 708 Right aagccatcttcaaagttgcagta 5701 Left gactccaagcacaccatcct 5702 Right agccatcttcaaagttgcagtaaaa b 709 Left agatggacttgctggatggg b 710 Right cattccaacaagtgaaggagctc 5703 Left gctectnctccactcaacac 5704 Right atctggctctcatatctccctg b 711 Left gaggctectactecttctcaa b 712 Ri ht ctctcatcttctccctgggc 5725 Left cagagctggtcctggcg 5713 Left tnctcatctcaacacctcagct 5726 Right aacaagtgaaggagctctgca 5714 Right gcagtttccggcacatctg 5727 Left cagatggacttgctggat 5715 Left ccgcatcccctccgag 5728 Right gactgtcc cattccaacaagtg 5716 Right cccattccaacaagtgaaggag 5729 Left ccatcctgagtccgtggat 5717 Left cctgagtccgtggatgagg 5730 Right gtccagccacagaagcc 5718 Right tccagccacagaagccatc 5731 Left ccgtggatgaggagcagc Seq. 5732 Right tgtgccttgggtccagc ID Primer Sequence 5719 Left ccagaggctectactccttc 5733 Left ctcagctgactccaagcaca 5720 Right ggaagtcacaggcctgcc 5734 Right gcacatctggctctcatc 5721 Left ttctcaacacctcagctgactccaa 5735 Left tcctggcgccgcatc 5722 Right cacaggcagtttccggc 5736 Right tgaaggagctctgcaggg 5723 Left tccaagcacaccatcctgag 5737 Left gctgactccaagcacaccat 5724 Right aagccatchcaaagttgca 5738 Right gcagthccggcacat ALK Region4 476-525 bases Seq. 5758 Right tgacagtgtgccttgggtc ID Primer Sequence 5739 Left ctectnctecttctcaacacct 5759 Left ccagaggctectnctecttc 5740 Right agccatcttcaaagttgcagtaaaa 5760 Right gcagtttc cggcacatctg Seq.
Primer Sequence ID 5761 Left aacacctcagctgactccaag 5741 Left gaggctectnctecttctcaac 5762 Right agtgtgccttgggtccag 5742 Right aagccatchcaaagttgcagta ___________________________________________ 5763 Left ctccaagcacaccatcctga 5743 Left gagtattccc ctc cactgcat 5764 Right agtgtggggtgacagtgtg 5744 Right cattccaacaagtgaaggagctc ___________________________________________ 5765 Left gtattcccctccactgcatgacctc 5745 Left tattcc cctc cactgcatgac 5766 Right tgaaggagctctgcaggg 5746 Right cccattccaacaagtgaaggag ___________________________________________ 5767 Left acaccatcctgagtccgtg 5747 Left agatggacttgctggatggg 5768 Right gtcctgacctgccattgag 5748 Right acatctggctctcatcttctcc ___________________________________________ 5769 Left ctcagctgactccaagcaca 5749 Left cactgcatgacctcaggaac 5770 Right ggggtgacagtgtgcctt 5750 Right gactgtc ccattc caacaagtg ___________________________________________ 5771 Left cccagaggctectnctcc 5751 Left gcatgacctcaggaaccaga 5772 Right aagccatcttcaaagttgca 5752 Right caacaagtgaaggagctctgc ___________________________________________ 5773 Left agagcgttctaaggagatg 5753 Left cactcaacacctcagctgactc 5774 Right gcacatctggctctcatcttc 5754 Right tccagccacagaagccatc ___________________________________________ 5775 Left accatcctgagtccgtggat 5755 Left ctgactccaagcacaccatc 5776 Right gtccagccacagaagcc 5756 Right attgaggagtgtggggtgac ___________________________________________ 5777 Left tccactgcatgacctcagg 5757 Left ctcatctcaacacctcagct 5778 Right gactgtcc cattccaacaa ALK Region4 750-1250 bases Seq.
Primer Sequence ID 5799 Left cctgattattttacatggaatctcacc 5779 Left tggaatctcacctggataatgaaag 5800 Right tgtcagacacatcgaggagag 5780 Right ttngttctccactagcaccaag _________________________________________ _5801 Left ctectnctccttctcaacacctc 5781 Left gaatcaccaacaaacatgccttc 5802 Rjght ctecttcccggtittgactc 5782 Right agccatcttcaaagttgcagtaaaa 5803 Left gctectnctccttctcaacac 5783 Left cacctggataatgaaagactccttc 5804 Ri ht ttt tctccactagcaccaaggac 5784 Right tggtcactgtagcactttcagaa 5805 Left aaa actcatcccMcct t 5785 Left tggataatgaaagactc cttc cctt _5806 Right ctcaagactccacgaatgagc 5786 Right caatagagcatggtcttggtgg 5787 Left attttacatggaatctcac ctggat 5788 Right gaaacgtagcactggtcactgtag 5807 Left aatcaccaacaaacatgccttctcc ___________________________________________ 5808 Right ctagcaccaaggacacgtttc 5789 Left atctcacctggataatgaaagactc 5790 Right cacccggtittgactccactag 5809 Left effete cactectgattatntaca ___________________________________________ 5810 Right cttcccggtittgactccac 5791 Left tccttctc ctgattattttacatgga 5792 Right aagccatchcaaagttgcagta 5811 Left tctectgattatittacatggaatctc 5812 Right gtagcactggtcactgtagcacttt Seq. 5813 Left gactggtcatagctc cttgga Primer Sequence ID 5814 Right gcacatctggctctcatcttc 5793 Left acatggaatctcacctggataatga 5794 Right gaaacgtagcactggtcactg 5815 Left gagaagaaggcgtcggaagt ___________________________________________ 5816 Right cattccaacaagtgaaggagctc 5795 Left gactggtcatagctccttggaatc 5796 Right gghttgactccactagcacc 5817 Left gatatcgggactggtcatagctc ___________________________________________ 5818 Right atctggctctcatcttctccctg 5797 Left cagatcttcgggactggtcatag 5798 Right acatctggctctcatcttctcc Table 12. EGFR cDNA Capture Primer List for NGS Panel EGFR Regionl 75-125 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
5819 Left CCCAGTGGAGAAGCTC 5823 Left CCCAGTGGAGAAG
5820 Right GGGATCCAGAGTCCCTTATACA 5824 Right TGGGATCCAGAGTCCC
5821 Left GAGAAGCTCCCAACCA 5825 Left GAGAAGCTCCCAA
5822 Right GGGATCCAGAGTCCCTTAT 5826 Right TGGGATCCAGAGT
EGFR Regionl 126-175 bases Seq. 5827 Left TTGTGGAGCCTCTTACACCC
Primer Sequence ID f, 828 Right ACGGGAATTTTAACTTTCTCACCTT

f841 Left TGCTGCAGGAGAGGGA
5829 Left CTTGTGGAGCCTCTTACACCCAGT _ 842 Right GGGATCCAGAGTCCCTTAT
5830 Right TGATAGCGACGGGAATTTTAACT
f843 Left CTTGTGGAGCCTCTT
5831 Left GGAGAGGGAGCTTGTGGAG f844 Right CCTTGATAGCGACGGGAATTTTA

5832 Right GGGATCCAGAGTCCCTTATACA
f845 Left GAGAAGCTCCCAA
5833 Left CTTGTGGAGCCTCTTACA f846 Right GCGACGGGAATTTTAACTTTCT
5834 Right CGACGGGAATTTTAACTTTCTCAC
f847 Left GCACGCTGCGGA
5835 Left CTGCAGGAGAGGGAGCTTG 5848 Right TGGGATCCAGAGTCCC
5836 Right AATTTTAACTTTCTCACCTTCTG
5849 Left CCCAGTGGAGAAG
5837 Left CCCAGTGGAGAAGCTC 5850 Right TCTCTTAATTCCTTGATAGCG
5838 Right TTCTCTTAATTCCTTGATAGCGACG
5851 Left CGGAGGCTGCTG
5839 Left GAGAAGCTCCCAACCA f 852 Right TGGGATCCAGAGT
5840 Right CTTAATTCCTTGATAGCGACGGGAA
Seq. f 853 Left TGCTGCAGGAGAG
Primer Sequence ID 4854 Right CTTCTGGGATCCA
EGFR Regionl 176-225 bases Sea 5872 Right TCCTTGATAGCGACGGGAATTT
ID.' Primer Sequence 5855 Left TTGTGGAGCCTCTTACACCC f 873 Left GCGCCACATCGTTCGGAA
5856 Right TTCTCTTAATTCCTTGATAGCGACG f, 874 Right AATTTTAACTTTCTCACCTTCTG
5875 Left GGATCGGCCTCTTCATGC
Sea 5876 Right GGGATCCAGAGTCCCTTAT
ID.' Primer Sequence 5857 Left CAGGAGAGGGAGCTTGTGG f 877 Left CCCAGTGGAGAAGCTC
5858 Right ACGGGAATTTTAACTTTCTCACCTT f, 878 Right ACGTAGGCTTCATCGAGGATTT
5879 Left CTTGTGGAGCCTCTTACA
5859 Left CTGCAGGAGAGGGAGCTTG f880 Right CCTTGTTGGCTTTCGGAGA
5860 Right TGATAGCGACGGGAATTTTAACTT _____ f881 Left GAGAAGCTCCCAACCA
5861 Left CTTGTGGAGCCTCTTACACCCAGT 7 882 Right CACGTAGGCTTCATCGAGGA
5862 Right CTTAATTCCTTGATAGCGACGGGAA _____ 7 883 Left CGAAGGCGCCACATCG
5863 Left CCACATCGTTCGGAAGCG f884 Right TGGGATCCAGAGTCCC
_ 5864 Right CGACGGGAATTTTAACTTTCTCAC ______ _________________________________________ 7 885 Left TGCTGCAGGAGAGG
5865 Left CATCGTTCGGAAGCGCAC f886 Right AGATGTTGCTTCTCTTAATTCC
_ 5866 Right CCTTGATAGCGACGGGAATTTTAA
__________________________________________ 5887 Left GCACGCTGCGGA
5867 Left ATCGGCCTCTTCATGCGAA 7888 Right TTCTCTTAATTCCTTGATAGCG
_ 5868 Right GGGATCCAGAGTCCCTTATACA
_________________________________________ 7889 Left CCCAGTGGAGAAG
5869 Left GAAGGCGCCACATCGTTC 7890 Right GGCCATCACGTAGGCTTCAT
5870 Right GCGACGGGAATTTTAACTTTCT
7891 Left CTTGTGGAGCCTCTT
_ 5871 Left TGCTGCAGGAGAGGGAG f, 892 Right TCGGAGATGTTGCTTCT

5894 Right CTGGCCATCACGTAGGCTT
5893 Left GAGAAGCTCC CAA
EGFR Regionl 226-275 bases S eq.
Primer Sequence ID _ 915 Left CTGGTGGTGGCCCTGG
5895 Left ATCGGCCTCTTCATGCGAA f916 Right TTCCTTGATAGCGACGGGAATTT

5896 Right TTCTCTTAATTCCTTGATAGCGACG
f917 Left CTGCAGGAGAGGGAGCTTG
5897 Left CCTCCTCTTGCTGCTGGT _ 918 Right CTGGCCATCACGTAGGCTT
5898 Right ACGGGAATTTTAACTTTCTCACCTT
__________________________________________ f919 Left CTGCTGGTGGTGGCCC
5899 Left TTGTGGAGCCTCTTACACCC f920 Right GCGACGGGAATTTTAACTTTCT
5900 Right CGTAGGCTTCATCGAGGATTTC
f921 Left CATCGTTCGGAAGCGCAC
5901 Left GAAGGCGCCACATCGTTC f922 Right CCTTGTTGGCTTTCGGAGA
5902 Right TGATAGCGACGGGAATTTTAACTT _____ 7 923 Left GCGCCACATCGTTCGGA
5903 Left CCTCTTGCTGCTGGTGGT f924 Right AGATGTTGCTTCTCTTAATTCC
5904 Right CGACGGGAATTTTAACTTTCTCAC
f925 Left TGCTGCAGGAGAGGGAG
5905 Left CCACATCGTTCGGAAGCG f926 Right TCACGTAGGCTTCATCGAG
5906 Right CTTAATTCCTTGATAGCGACGGGAA
f927 Left CGAAGGCGCCACATCG
5907 Left GGATCGGCCTCTTCATGC f928 Right TTCTCTTAATTCCTTGATAGCG
5908 Right CCTTGATAGCGACGGGAATTTTAA
f929 Left GGGCCCTCCTCTTGCT
5909 Left GGAGAGGGAGCTTGTGGAG f930 Right GGGATCCAGAGTCCCTTAT
5910 Right CACGTAGGCTTCATCGAGGAT
S eq. _ 931 Left CTTGTGGAGCCTCTTACA
Primer Sequence ID _ 932 Right CGCTGGCCATCACGTAGG
5911 Left GCCCTCCTCTTGCTGCT
5912 Right GGGATCCAGAGTCCCTTATACA f933 Left GAGAAGCTCCCAAC CA
5934 Right GGTGGAGGTGAGGCAGATG
5913 Left CTTGTGGAGCCTCTTACACCCAGT
5914 Right GGCCATCACGTAGGCTTCAT
EGFR Region2 75-125 bases Seq.
Primer Sequence Primer Sequence ID ID
5935 Left AATCCTCGATGAAGCCTACGT 5939 Left GGAAATCCTCGATGAAGCCTA
5936 Right CCAGGAGGCAGCCGAAG 5940 Right CGAAGGGCATGAG
5937 Left AATCCTCGATGAAGCCTACGTGATG
5938 Right AGGAGGCAGCCG
EGFR Region2 126-175 bases S ea 944 Right AATATTGTCTTTGTGTTCCCGGAC
ID.' Primer Sequence 5941 Left GAAATCCTCGATGAAGCCTACG 5945 Left AACATCTCCGAAAGCCAACAAG
5942 Right GAGCCAATATTGTCTTTGTGTTCC 5946 Right CTTTGTGTTCCCGGACATAGTC
5943 Left GAAATCCTCGATGAAGCCTACGTGA 5947 Left CTCGATGAAGCCTACGTGATGG

5948 Right ATTGTCTTTGTGTTCCCGGACATA 5956 Right AGGAGGCAGCCG
5949 Left TCGCTATCAAGGAATTAAGAGAAGC 5957 Left TTAAAATTCCCGTCGCTATCAAGG
5950 Right CCAGGAGGCAGCCGAAG 5958 Right CGAAGGGCATGAG
5951 Left GGAAATCCTCGATGAAGCC 5959 Left GGAAATCCTCGATGAA
5952 Right GGGAGCCAATATTGTCTTTGTGT 5960 Right TGGGAGCCAATATTGTCTTTG
Seq.
Primer Sequence ID 961 Left CAACAAGGAAATCC
5953 Left CAAGGAATTAAGAGAAGCAACATCT _ 962 Right TGGGAGCCAATATTGTCT
5954 Right GACATAGTCCAGGAGG
5955 Left CCGTCGCTATCAAGGAATTAAGAG
¨
EGFR Region2 176-225 bases Seq.
Primer Sequence ID 5983 Left TTAAAATTCCCGTCGCTATCAAGG
5963 Left GAAAGTTAAAATTCCCGTCGCTATC 7984 Right CCAGGAGGCAGCCGAAG
5964 Right GAGCCAATATTGTCTTTGTGTTCC
7985 Left GAAGCAACATCTCCGAAAGCCAA
5965 Left GAAGGTGAGAAAGTTAAAATTCCCG 5986 Right CGATCTGCACACACCAGTT
5966 Right ATTGTCTTTGTGTTCCCGGACATAG
5987 Left AATTCCCGTCGCTATCAAGGAATTA
5967 Left GTGAGAAAGTTAAAATTCCCGTCG 5988 Right TGGGAGCCAATATTGTCT
5968 Right AATATTGTCTTTGTGTTCCCGGAC
5989 Left AAATCCTCGATGAAGCCT
5969 Left CCGTCGCTATCAAGGAATTAAGAG 5990 Right CCTCCAAGTAGTTCATGCCCTTT
5970 Right GGGAGCCAATATTGTCTTTGTGT
5991 Left TCAAGGAATTAAGAGAAGCAACATCT
5971 Left GAAATCCTCGATGAAGCCTACG 5992 Right GACATAGTCCAGGAGG
5972 Right GACGGTCCTCCAAGTAGTTCAT
Seq. 5993 Left TATCAAGGAATTAAGAGAAGCAACA

Primer Sequence ID 5994 Right GGTACTGGGAGCCA
5973 Left TCGCTATCAAGGAATTAAGAGAAGC _______ 5974 Right TGGGAGCCAATATTGTCTTTG 5995 Left CCCAGAAGGTGAGAAAGTTAAAAT
5996 Right AGGAGGCAGCCG
5975 Left AGAAGCAACATCTCCGAAAGC
5976 Right GATCTGCACACACCAGTTGAG 5997 Left AGAAGCAACATCTCCGAA
5998 Right CGATCTGCACACACCA
5977 Left GAAATCCTCGATGAAGCCTACGTGA _______ 5978 Right CGACGGTCCTCCAAGTAGTT 5999 Left GAAATCCTCGATGAAG
6000 Right GCGACGGTCCTCCAAGTA
5979 Left CTCGATGAAGCCTACGTGATGG
5980 Right TCCTCCAAGTAGTTCATGCCC 6001 Left GGCACGGTGTATAAGGGAC
6002 Right CGAAGGGCATGAG
5981 Left CAACATCTCCGAAAGCCAACAA
5982 Right CACACACCAGTTGAGCAGG
EGFR Region2 226-275 bases Seq. 6005 Left GAAAGTTAAAATTCCCGTCGCTATC

Primer Sequence ID 6006 Right TGGGAGCCAATATTGTCTTTGTG
6003 Left AAGGTGAGAAAGTTAAAATTCCCGT
6004 Right GGAGCCAATATTGTCTTTGTGTTC E007 Left TCGCTATCAAGGAATTAAGAGAAGC
E008 Right GACGGTCCTCCAAGTAGTTCAT

6025 Left CCCAGAAGGTGAGAAAGTTAAAAT
6009 Left GTGAGAAAGTTAAAATTCCCGTCGC 6026 Right GTCTTTGTGTTCCCGGACATA
6010 Right CCAATATTGTCTTTGTGTTCCCG
6027 Left AATTCCCGTCGCTATCAAGGAAT
6011 Left CCGTCGCTATCAAGGAATTAAGAG 6028 Right CGATCTGCACACACCAGTT
6012 Right CCTCCAAGTAGTTCATGCCCTTT
6029 Left AACATCTCCGAAAGCCAACAAG
6013 Left TCAAGGAATTAAGAGAAGCAACATCT 6030 Right AAGCGACGGTCCTCCAAGTA
6014 Right TCCTCCAAGTAGTTCATGCCC
6031 Left GGCACGGTGTATAAGGGAC
6015 Left TTAAAATTCCCGTCGCTATCAAGG 6032 Right CTTTGTGTTCCCGGACATAGTC
6016 Right GATCTGCACACACCAGTTGAG
6033 Left TCGATGAAGCCTACGTGATGG
6017 Left GAAATCCTCGATGAAGCCTACG 6034 Right GACATGCTGCGGTGTTTTCA
6018 Right TGTTTTCACCAGTACGTTCCTG
6035 Left GAAATCCTCGATGAAGCCTACGTGA
6019 Left AGAAGGTGAGAAAGTTAAAATTCC 6036 Right AGTACGTTCCTGGCTGCC
6020 Right ATATTGTCTTTGTGTTCCCGGAC
6037 Left AGAAGCAACATCTCCGAAAGC
6021 Left TATCAAGGAATTAAGAGAAGCAACA 6038 Right CAAGCGACGGTCCTCCAA
6022 Right CGACGGTCCTCCAAGTAGTT
6039 Left AAAAGATCAAAGTGCTGGGCTC
Seq. 6040 Right CCAGGAGGCAGCCGAAG
ID Primer Sequence 6023 Left ATTCCCGTCGCTATCAAGGAATTAA 6041 Left GAAGCAACATCTCCGAAAGCCAAC
6024 Right CACACACCAGTTGAGCAGG 6042 Right CTGCCAGGTCGCGGT
EGFR Region3 75-125 bases S ea 045 Left AAAGGGCATGAACTACTTGGAGGAC

ID' Primer Sequence _ 046 Right CACCCAGCAGTTTG
6043 Left CAAAGGGCATGAACTACTTGGAG 6047 Left GCATGAACTACTTG
6044 Right CCAGCAGTTTGGCC 6048 Right TTCCGCACCCAG
Seq.
Primer Sequence ID
EGFR Region3 126-175 bases S eq. 059 Left AAAGGGCATGAACTACTTG
Primer Sequence ID 060 Right GC CTCCTTCTGCATGGTATTCT
6049 Left CAAAGGGCATGAACTACTTGGAG Seq.
6050 Right CCTTCTGCATGGTATTCTTTCTCTT ID Primer Sequence 061 Left GGCTCCCAGTACCTGCTC
6051 Left AAAGGGCATGAACTACTTGGAGGAC 6062 Right CCAGCAGTTTGGCC
6052 Right CCTCCTTCTGCATGGTATTCTTTC
063 Left CAAAGGGCATGAACTAC
6053 Left CAACTGGTGTGTGCAGATCG 6064 Right CACTTTGCCTCCTTCTGC
6054 Right CTGCATGGTATTCTTTCTCTTCCG
065 Left GCTCAACTGGTGTGTGCAGA
6055 Left CTCCCAGTACCTGCTCAACT 6066 Right CACCCAGCAGTTTG
6056 Right CTTTCTCTTCCGCACCCAG
067 Left CAGATCGCAAAGGG
6057 Left GCTCAACTGGTGTGTGC 6068 Right GC CTCCTTCTGCATGGTAT
6058 Right CATGGTATTCTTTCTCTTCCGCAC
069 Left CAAAGGGCATGAA

6070 Right GCCTCCTTCTGCATGG
EGFR Region3 176-225 bases S eq. . 6090 Right TTTCTCTTCCGCACCCAG
ID Pnmer Sequence 6071 Left CAAAGGGCATGAACTACTTGGAG 6091 Left CAAAGGGCATGAACTACTTG
6072 Right CTCTGGTGGGTATAGATTCTGTGTA 6092 Right AGATTCTGTGTAAAATTGATTCCA
6073 Left CTATGTCCGGGAACACAAAGAC 6093 Left TGGCTCCCAGTACCTG
6074 Right CCTTCTGCATGGTATTCTTTCTCTT 6094 Right CACTTTGCCTCCTTCTGC
6075 Left GACTATGTCCGGGAACACAAA 6095 Left GACTATGTCCGGGAACAC
6076 Right CCTCCTTCTGCATGGTATTCTTTC 6096 Right GCCTCCTTCTGCATGG
6077 Left AAAGGGCATGAACTACTTGGAGGAC 6097 Left CTTCGGCTGCCTCCTGG
6078 Right ACTCTGGTGGGTATAGATTCTGT 6098 Right CCAGCAGTTTGGCC
6079 Left CTCCCAGTACCTGCTCAACT 6099 Left AAAGGGCATGAACTAC
6080 Right CTGCATGGTATTCTTTCTCTTCCG 6100 Right ACTCTGGTGGGTATAGATTC
6081 Left CTCAACTGGTGTGTGCAGATC 6101 Left CTGCTCAACTGGTGT
6082 Right GCCTCCTTCTGCATGGTATTCT 6102 Right TTCCAATGCCATCCACTT
6083 Left GGCTCCCAGTACCTGCTC 6103 Left CCCTTCGGCTGCCTCC
6084 Right CATGGTATTCTTTCTCTTCCGCAC 6104 Right CACCCAGCAGTTTG
6085 Left GTCCGGGAACACAAAGACAATATT 6105 Left ACAATATTGGCTCCCA
6086 Right GCCTCCTTCTGCATGGTAT 6106 Right ATCCACTTGATAGGCAC
6087 Left CTGCTCAACTGGTGTGTGC 6107 Left CAGATCGCAAAGGG
6088 Right TTCCAATGCCATCCACTTGAT 6108 Right AGATTCTGTGTAAAATTGAT
S eq. . 6109 Left CAAAGGGCATGAAC
Pnmer Sequence ID 6110 Right CTCTGGTGGGTATAGA
6089 Left GTCCGGGAACACAAAGACAAT
EGFR Region3 226-275 bases S eq. . 6120 Right CCTCCTTCTGCATGGTATTCTTTC

ID Pnmer Sequence 6111 Left GTCCGGGAACACAAAGACAATATT 6121 Left CTCCACCGTGCAGCTCAT
6112 Right ATTCCAATGCCATCCACTTGAT 6122 Right CTGCATGGTATTCTTTCTCTTCCG

6113 Left CTCCCAGTACCTGCTCAACT 6123 Left CGTGCAGCTCATCACGC
6114 Right CTCTGGTGGGTATAGATTCTGTGTA 6124 Right CATGGTATTCTTTCTCTTCCGCAC
6115 Left GCTCATCACGCAGCTCATG 6125 Left CTATGTCCGGGAACACAAAGAC
6116 Right CCTTCTGCATGGTATTCTTTCTCTT 6126 Right GATTCTGTGTAAAATTGATTCCA
6117 Left CAACTGGTGTGTGCAGATCG 6127 Left GCTCAACTGGTGTGTGCAG
6118 Right ACTCTGGTGGGTATAGATTCTGT 6128 Right CCGTAGCTCCAGACATCACT
6119 Left CAGCTCATCACGCAGCTC 6129 Left CTATGTCCGGGAACACAAAGACAAT

6130 Right CACTTTGCCTCCTTCTGC 6140 Right CCCGTAGCTCCAGACATC
S eq. 6141 Left CCTCCACCGTGCAGCT
Primer Sequence ID 6142 Right GC CTCCTTCTGCATGGTAT
6131 Left CTTCGGCTGCCTCCTGG
¨
6132 Right GCCTCCTTCTGCATGGTATTCT 6143 Left TGGCTCCCAGTACCTGC
6144 Right ACTCTGGTGGGTATAGATTC
6133 Left CTGCCTCACCTCCACCG
6134 Right CTTTCTCTTCCGCACCCAG 6145 Left CCCTTCGGCTGCCTCC
6146 Right GCCTCCTTCTGCATGG
6135 Left GACTATGTCCGGGAACACAAA
6136 Right ATTCCAATGCCATCCACTT 6147 Left CAGATCGCAAAGGG
6148 Right AAAGGTCATCAACTCCCAAACAG
6137 Left CTGCTCAACTGGTGTGTG
6138 Right AAACAGTCACCCCGTAGCTC 6149 Left CTGCTCAACTGGTGT
6150 Right CACCCCGTAGCTCCAGAC
6139 Left GCTCCCAGTACCTGCTCA
EGFR Region4 276-325 bases Sea Seq.
Primer Sequence Primer Sequence ID
6151 Left CTCCCAACCAAGCTCTCTT 6159 Left TTGTGGAGCCTCTTACACCC
6152 Right TTGTGTTCCCGGACATAGTC 6160 Right CGAAGGGCATGAG
6153 Left GAGAAGCTCCCAACCAAG 6161 Left CC CAGTGGAGAAGCTC
6154 Right CCAGGAGGCAGCCGAAG 6162 Right AGGAGGCAGCCG
6155 Left TCTCTTGAGGATCTTGA 6163 Left GAGAAGCTCCCAACC
6156 Right GCCAATATTGTCTTTGTGTTCCC 6164 Right GACATAGTCCAGGAGG
6157 Left CTCCCAACCAAGCTCT
6158 Right TTGTGTTCCCGGACATA
EGFR Region4 326-375 bases Sea ID.' Primer Sequence 6177 Left CTCCCAACCAAGCTCT
6165 Left CTTGTGGAGCCTCTTACACCCAG 6178 Right GATCTGCACACACCAGTTGAG
6166 Right GAGCCAATATTGTCTTTGTGTTCC
6179 Left TCTCTTGAGGATCTTGA
6167 Left AGGAGAGGGAGCTTGTGGA 6180 Right CGATCTGCACACACCAGTT
6168 Right ATTGTCTTTGTGTTCCCGGACATAG Seq.
ID Primer Sequence 6169 Left CTCCCAACCAAGCTCTCTT 6181 Left CTTGTGGAGCCTCTTAC
6170 Right AATATTGTCTTTGTGTTCCCGGAC 6182 Right TGGGAGCCAATATTGTCTT
6171 Left CTTGTGGAGCCTCTTACACC 6183 Left TGCTGCAGGAGAGGGAG
6172 Right TGGGAGCCAATATTGTCTTTGT 6184 Right GACATAGTCCAGGAGG
6173 Left CTGCAGGAGAGGGAGCTTG 6185 Left CCACATCGTTCGGAAGCG
6174 Right CCAGGAGGCAGCCGAAG 6186 Right CGAAGGGCATGAG
6175 Left GAGAAGCTCCCAACCAAG 6187 Left CC CAGTGGAGAAGCT
6176 Right CACACACCAGTTGAGCAGG 6188 Right CCAGTTGAGCAGGTAC

6189 Left GAGAAGCTCCCAACC 6193 Left TGTGGAGCCTCT
6190 Right TGGGAGCCAATATTGT 6194 Right TACTGGGAGCCA
6191 Left TCGGAAGCGCACG
6192 Right AGGAGGCAGCCG
EGFR Region4 376-425 bases Seq. .
Pnmer Sequence ID 6215 Left GAGAAGCTCCCAACCAAG
6195 Left AGGAGAGGGAGCTTGTGGA 6216 Right CGACGGTCCTCCAAGTAGTT
6196 Right GAGCCAATATTGTCTTTGTGTTCC _____ 6217 Left CTTGTGGAGCCTCTTACA
6197 Left GAAGGCGCCACATCGTTC 6218 Right CACACACCAGTTGAGCAGG
6198 Right AATATTGTCTTTGTGTTCCCGGAC
6219 Left CCCAGTGGAGAAGCTC
6199 Left TTGTGGAGCCTCTTACACCC _6220 Right AAGCGACGGTCCTCCAAGTA
6200 Right CTCCAAGTAGTTCATGCCCTTT
_6221 Left CGCCACATCGTTCGGAA
6201 Left CCACATCGTTCGGAAGCG 6222 Right TGGGAGCCAATATTGTCT
6202 Right GGGAGCCAATATTGTCTTTGTGT
6223 Left CTCCCAACCAAGCTCT
6203 Left CTGCAGGAGAGGGAGCTTG 6224 Right CAAGCGACGGTCCTCCAA
6204 Right TTGTCTTTGTGTTCCCGGACATAG ______ 6225 Left GAGAAGCTCCCAACC
6205 Left TGGAGCCTCTTACACCCAGT 6226 Right CGATCTGCACACACCAGTT
6206 Right TCCTCCAAGTAGTTCATGCCC
6227 Left TCTCTTGAGGATCTTGA
6207 Left CTCCCAACCAAGCTCTCTT 6228 Right GTACGTTCCTGGCTGCCA
6208 Right GACGGTCCTCCAAGTAGTTCAT
6229 Left GGATCGGCCTCTTCATGC
6209 Left TGCTGCAGGAGAGGGAG 6230 Right GACATAGTCCAGGAGG
6210 Right GATCTGCACACACCAGTTGAG
Seq. . 6231 Left CTGGTGGTGGCCCTGG
Pnmer Sequence ID 6232 Right AGGAGGCAGCCG
6211 Left CATCGTTCGGAAGCGCAC
6212 Right TGGGAGCCAATATTGTCTTTG 6233 Left CCTCTTGCTGCTGGTGGT
___________________________________________ 6234 Right CGAAGGGCATGAG
6213 Left ATCGGCCTCTTCATGCGAA
6214 Right CCAGGAGGCAGCCGAAG
EGFR Region4 426-475 bases Seq. . 6242 Right GGGAGCCAATATTGTCTTTGTGT

ID Pnmer Sequence 6235 Left ATCGGCCTCTTCATGCGAA 6243 Left CTTGTGGAGCCTCTTACACCCAGT

6236 Right GAGCCAATATTGTCTTTGTGTTCC 6244 Right CCTCCAAGTAGTTCATGCCCTTT
6237 Left CCTCCTCTTGCTGCTGGT 6245 Left CCTCTTGCTGCTGGTGGT
6238 Right AATATTGTCTTTGTGTTCCCGGAC 6246 Right ATTGTCTTTGTGTTCCCGGACATAG
6239 Left TTGTGGAGCCTCTTACACCC 6247 Left GGAGAGGGAGCTTGTGGAG
6240 Right GACGGTCCTCCAAGTAGTTCAT 6248 Right TCCTCCAAGTAGTTCATGC CC
6241 Left GAAGGCGCCACATCGTTC 6249 Left CCACATCGTTCGGAAGCG

6250 Right GATCTGCACACACCAGTTGAG 6262 Right AAGCGACGGTCCTCCAAGTA
6251 Left CTCCCAACCAAGCTCTCTT 6263 Left GCCCTCCTCTTGCTGCT
6252 Right TGTTTTCACCAGTACGTTCCTG 6264 Right CCAGGAGGCAGCCGAAG
6253 Left CTGCAGGAGAGGGAGCTTG 6265 Left CTTGTGGAGCCTCTTACA
6254 Right CGACGGTCCTCCAAGTAGTT 6266 Right AGTACGTTCCTGGCTGCC
S eq. 6267 Left TCTCTTGAGGATCTTGA
Primer Sequence ID 6268 Right CCAAAATCTGTGATCTTGACATGC
6255 Left CATCGTTCGGAAGCGCAC
6256 Right CACACACCAGTTGAGCAGG 6269 Left GAAGGCGCCACATCG
6270 Right CGATCTGCACACACCAGTT
6257 Left GAGAAGCTCCCAACCAAG
6258 Right GATCTTGACATGCTGCGGTGTTTTC 6271 Left CTCCCAACCAAGCTCT
6272 Right GATCTTGACATGCTGCGGTG
6259 Left GCGCCACATCGTTCGGAA
6260 Right TGGGAGCCAATATTGTCTTTG 6273 Left CTGGTGGTGGCCCTGG
6274 Right TGGGAGCCAATATTGTCT
6261 Left TGCTGCAGGAGAGGGAG
EGFR Region5 276-325 bases Seaeq.
Primer Sequence Primer Sequence ID
6275 Left ATCCTCGATGAAGCCTACGTG 6279 Left GGAAATCCTCGATGAAGCCTAC
6276 Right TCTCTTCCGCACCCAG 6280 Right CCAGCAGTTTGGCC
6277 Left CGATGAAGCCTACGTGATGG 6281 Left GGAAATCCTCGATGAAGCC
6278 Right TTCTTTCTCTTCCGCAC 6282 Right CACCCAGCAGTTTG
EGFR Region5 326-375 bases S eq.
Primer Sequence ID 6297 Left CTCGATGAAGCCTACGTGATGG
6283 Left GAAATCCTCGATGAAGCCTACG 6298 Right CACTTTGCCTCCTTCTGC
6284 Right CCTTCTGCATGGTATTCTTTCTCTT _ S eq.
TCAAGGAATTAAGAGAAGCAACATC
Primer Sequence ID 6299 Left T
6285 Left GAAATCCTCGATGAAGCCTACGTGA 6300 Right CCAGCAGTTTGGCC
6286 Right CTGCATGGTATTCTTTCTCTTCCG
6301 Left CCGTCGCTATCAAGGAATTAAGAG
6287 Left AAGCAACATCTCCGAAAGC CAA 6302 Right CACCCAGCAGTTTG
6288 Right CTCCTTCTGCATGGTATTCTTTCT
6303 Left GGAAATCCTCGATGAAG
6289 Left AACATCTCCGAAAGCCAACAAG 6304 Right GCCTCCTTCTGCATGGT
6290 Right GCCTCCTTCTGCATGGTATT
__________________________________________ 6305 Left GGAAATCCTCGATG
6291 Left AGAAGCAACATCTCCGAAAG 6306 Right ATCCACTTGATAGGCAC
6292 Right CATGGTATTCTTTCTCTTCCGCAC
¨6307 Left CAACAAGGAAATCC
6293 Left GGAAATCCTCGATGAAGCCT 6308 Right CACTTTGCCTCCTTC
6294 Right GCCTCCTTCTGCATGGTATTCTT
6295 Left TCGCTATCAAGGAATTAAGAGAAGC _ _________________________________________________ 6296 Right TCTCTTCCGCACCCAG

EGFR Region5 376-425 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
6309 Left AAGGTGAGAAAGTTAAAATTCCCGT 6329 Left GCAACATCTCCGAAAGCCAA
6310 Right CCTTCTGCATGGTATTCTTTCTCTT 6330 Right AGATTCTGTGTAAAATTGATTCCA
6311 Left GAAAGTTAAAATTCCCGTCGCTATC 6331 Left TCAAGGAATTAAGAGAAGCAACATCT
6312 Right CTCCTTCTGCATGGTATTCTTTCT 6332 Right CACTTTGCCTCCTTCTGC
6313 Left GAAATCCTCGATGAAGCCTACG 6333 Left AGAAGCAACATCTCCGAAAGC
6314 Right CTCTGGTGGGTATAGATTCTGTGTA 6334 Right ATTCCAATGCCATCCACTT
6315 Left CCGTCGCTATCAAGGAATTAAGAG 6335 Left ATTCCCGTCGCTATCAAGGAA
6316 Right CATGGTATTCTTTCTCTTCCGCAC 6336 Right GCCTCCTTCTGCATGGT
6317 Left GTTAAAATTCCCGTCGCTATCAAG 6337 Left TCGATGAAGCCTACGTGA
6318 Right TGCATGGTATTCTTTCTCTTCCG 6338 Right CCCGTAGCTCCAGACATCA
6319 Left ATTCCCGTCGCTATCAAGGAATTA 6339 Left GGAAATCCTCGATGAAGCCT
6320 Right GCCTCCTTCTGCATGGTATTCTT 6340 Right ACTCTGGTGGGTATAGATTC
6321 Left TCGCTATCAAGGAATTAAGAGAAGC 6341 Left CAGAAGGTGAGAAAGTTAAAATTCC
6322 Right GCCTCCTTCTGCATGGTATT 6342 Right CCAGCAGTTTGGCC
6323 Left AACATCTCCGAAAGCCAACAAG 6343 Left CCCAGAAGGTGAGAAAGTTAAAAT
6324 Right ATTCCAATGCCATCCACTTGAT 6344 Right CACCCAGCAGTTTG
6325 Left CTCGATGAAGCCTACGTGATGG 6345 Left TATCAAGGAATTAAGAGAAGCAACA

6326 Right ACTCTGGTGGGTATAGATTCTGT 6346 Right TCCAATGCCATC CA
6327 Left TGAGAAAGTTAAAATTCCCGTCGC 6347 Left AGAAGCAACATCTCCGAA
6328 Right CTTTCTCTTCCGCACCCAG 6348 Right ATCCACTTGATAGGCAC
EGFR Region5 426-475 bases Seq. 6360 Right CATGGTATTCTTTCTCTTCCGCAC

ID Primer Sequence 6349 Left AAGTTAAAATTCCCGTCGCTATCA 6361 Left TCAAGGAATTAAGAGAAGCAACATCT
6350 Right CTCTGGTGGGTATAGATTCTGTGTA 6362 Right CCCCGTAGCTCCAGACATC
6351 Left TCGCTATCAAGGAATTAAGAGAAGC 6363 Left GGCACGGTGTATAAGGGAC
6352 Right ATTCCAATGCCATCCACTTGAT 7364 Right CCTTCTGCATGGTATTCTTTCTCTT

6353 Left CCGTCGCTATCAAGGAATTAAGAG 6365 Left CCCAGAAGGTGAGAAAGTTAAA
6354 Right ACTCTGGTGGGTATAGATTCTGT 6366 Right CCTCCTTCTGCATGGTATTCTTTC

6355 Left AAGGTGAGAAAGTTAAAATTCCCGT 7367 Left AGAAGCAACATCTCCGAAAGC
6356 Right AGATTCTGTGTAAAATTGATTCCA 6368 Right CGTAGCTCCAGACATCACTCT
6357 Left GAAATCCTCGATGAAGCCTACG Sea 6358 Right GTCATCAACTCCCAAACAGTCAC ID ' Primer Sequence 6369 Left CTCGATGAAGCCTACGTGATGG
6359 Left AAAGATCAAAGTGCTGGGCTC 6370 Right TGGATCCAAAGGTCATCAACTC

6371 Left AACATCTCCGAAAGCCAACAAG 6381 Left GAAGCAACATCTCCGAAAGCCAAC

6372 Right CATCAACTCCCAAACAGTCACC CC 6382 Right CACCCCGTAGCTCCAGAC
6373 Left AAATCCTCGATGAAGCCTACGTGA 6383 Left TTAAAATTCCCGTCGCTATCAAGG
6374 Right AAACAGTCACCCCGTAGCTC 6384 Right ACTCTGGTGGGTATAGATTC
6375 Left AAAGATCAAAGTGCTGGGCTCCGG 6385 Left CAGAAGGTGAGAAAGTTAAAATTCC
6376 Right TGCATGGTATTCTTTCTCTTCCG 6386 Right CACTTTGCCTCCTTCTGC
6377 Left GGAAATCCTCGATGAAGCCT 6387 Left GGGCTCCGGTGCGTT
6378 Right AAGGTCATCAACTCCCAAACAG 6388 Right GC CTCCTTCTGCATGGTATTCT

6379 Left GAGAAAGTTAAAATTCCCGTCGCTA
6380 Right ATTCCAATGCCATCCACTT
EGFR Region6 476-525 bases S ea 400 Right GCCTCCTTCTGCATGGTATTC
ID.' Primer Sequence 6389 Left GGAGCCTCTTACACCCAGT 6401 Left TCTCTTGAGGATCTTGA
6390 Right TTCTGCATGGTATTCTTTCTCTTCC 6402 Right ATTCCAATGCCATCCACTTGAT
6391 Left CTCCCAACCAAGCTCTCTT 6403 Left CTCCCAACCAAGCTCT
6392 Right CTCCTTCTGCATGGTATTCTTTCTC ¨ 404 Right GCCTCCTTCTGCATGGTA
6393 Left CTTGTGGAGCCTCTTACAC 6405 Left CTGCAGGAGAGGGAGCTTG
6394 Right CATGGTATTCTTTCTCTTCCGCAC 6406 Right CCAGCAGTTTGGCC
6395 Left GAGAAGCTCCCAACCAAG 6407 Left GAGAAGCTCCCAACC
6396 Right CCTCCTTCTGCATGGTATTCTTT 6408 Right CACTTTGCCTCCTTCTGC
6397 Left GGAGAGGGAGCTTGTGGA
6398 Right TTTCTCTTCCGCACCCAG
6409 Left TGCTGCAGGAGAGGGA
__________________________________________ 6410 Right CACCCAGCAGTTTG
S ea ID.' Primer Sequence 6411 Left CCCAGTGGAGAAG
6399 Left CCCAGTGGAGAAGCTC 6412 Right GCCTCCTTCTGCATG
EGFR Region6 526-575 bases Sea 6421 Left CTCCCAACCAAGCTCTCTT
ID.' Primer Sequence 6422 Right ATTCCAATGCCATCCACTTGAT
6413 Left CTTGTGGAGCCTCTTACACCCAG
6414 Right CCTTCTGCATGGTATTCTTTCTCTT 6423 Left GAGAAGCTCCCAACCAAG
6424 Right GC CTC CTTCTGCATGGTATTCT
6415 Left CAGGAGAGGGAGCTTGTGG
6416 Right CTGCATGGTATTCTTTCTCTTCCG 6425 Left TGCAGGAGAGGGAGC
6426 Right CTTTCTCTTCCGCACCCAG
6417 Left CTGCAGGAGAGGGAGCTTG
¨
6418 Right CCTCCTTCTGCATGGTATTCTTTC 6427 Left CCCAGTGGAGAAGCTC
_6428 Right GCCTCCTTCTGCATGGTAT
6419 Left CTTGTGGAGCCTCTTACAC Sea 6420 Right CATGGTATTCTTTCTCTTCCGCAC ID'. Primer Sequence 6429 Left TCTCTTGAGGATCTTGA

6430 Right AGATTCTGTGTAAAATTGATTC CA 6437 Left CATCGTTCGGAAGCGCAC
6438 Right CACCCAGCAGTTTG
6431 Left CTCCCAACCAAGCTCT
6432 Right CACTTTGCCTCCTTCTGC 6439 Left CTTGTGGAGCCTCTTA
6440 Right GC CTC CTTCTGCATGG
6433 Left CCACATCGTTCGGAAGCG
6434 Right CCAGCAGTTTGGCC 6441 Left CCCAGTGGAGAAG
6442 Right ATCCACTTGATAGGCAC
6435 Left GAGAAGCTCCCAACC
6436 Right ATTCCAATGCCATCCACTT 6443 Left CTTGTGGAGCCTC
6444 Right CACTTTGCCTCCTTC
EGFR Region6 576-625 bases S eq.
Primer Sequence ID 6465 Left GAGAAGCTCCCAACCAAG
6445 Left CTTGTGGAGCCTCTTACACCCAG 6466 Right AGATTCTGTGTAAAATTGATTCCA
6446 Right CTCTGGTGGGTATAGATTCTGTGTA
6467 Left GCGCCACATCGTTCGGAA
6447 Left CCACATCGTTCGGAAGCG
6468 Right GC CTCCTTCTGCATGGTAT
6448 Right CCTTCTGCATGGTATTCTTTCTCTT
6469 Left CGAAGGCGCCACATCG
6449 Left ATCGGCCTCTTCATGCGAA 6470 Right CACTTTGCCTCCTTCTGC
6450 Right CATGGTATTCTTTCTCTTCCGCAC
¨6471 Left TGCTGCAGGAGAGGGAG
6451 Left GAAGGCGCCACATCGTTC 6472 Right ATTCCAATGCCATCCACTT
6452 Right CCTCCTTCTGCATGGTATTCTTTC
6473 Left CCCAGTGGAGAAGCTC
6453 Left CTTGTGGAGCCTCTTACACC 6474 Right CCCGTAGCTCCAGACATCAC
6454 Right ACTCTGGTGGGTATAGATTCTGT
6475 Left TCTCTTGAGGATCTTGA
6455 Left CAGGAGAGGGAGCTTGTGG 6476 Right AAAGGTCATCAACTCCCAAACAG
6456 Right ATTCCAATGCCATCCACTTGAT
6477 Left CTCCCAACCAAGCTCT
6457 Left CGTTCGGAAGCGCACG 6478 Right AAACAGTCACCCCGTAGCTC
6458 Right CTGCATGGTATTCTTTCTCTTCCG
S eq. 6479 Left GAGAAGCTCCCAACC
Primer Sequence ID 6480 Right AC CCCGTAGCTCCAGACAT
6459 Left CTGCAGGAGAGGGAGCTTG
6460 Right GCCTCCTTCTGCATGGTATTCT 6481 Left CTTGTGGAGCCTCTTAC
6482 Right ACTCTGGTGGGTATAGATTC
6461 Left CTCCCAACCAAGCTCTCTT
6462 Right CGTAGCTCCAGACATCACTCT 6483 Left CTCTTGCTGCTGGTGGTG
6484 Right CCAGCAGTTTGGCC
6463 Left GGATCGGCCTCTTCATGC
6464 Right CTTTCTCTTCCGCACCCAG
EGFR Region6 750-1250 bases Sea ID.' Primer Sequence 6489 Left ACCAAAATTATAAGCAACAGAGGTG
6485 Left TTTGTGGAGAACTCTGAGTGCATA 6490 Right CCTCCTTCTGCATGGTATTCTTT
6486 Right CTCTGGTGGGTATAGATTCTGTGTA
6491 Left TATAAGCAACAGAGGTGAAAACAGC
6487 Left CAGGAGTCATGGGAGAAAACAAC 6492 Right ACTCTGGTGGGTATAGATTCTGT
6488 Right CCTTCTGCATGGTATTCTTTCTCTT

6493 Left GAGTCATGGGAGAAAACAACACC
6494 Right TTTTGGAGAATTCGATGATCAACTC Left GTCAGAAAACCAAAATTATAAGCAA

6495 Left GAGTTTGTGGAGAACTCTGAGTG ¨6510 Right ATTCCAATGCCATCCACTTGAT
6496 Right CATGGTATTCTTTCTCTTCCGCAC _____ ¨6511 Left ATCCAAACTGCACCTACGGAT
6497 Left GAGAAAACAACACCCTGGTCTG 6512 Right GATCCAAAGGTCATCAACTCCCAA
6498 Right TGCATGGTATTCTTTCTCTTCCG
¨6513 Left CCATGAACATCACCTGCACAG
6499 Left AAAACAACACCCTGGTCTGGAAGTA 6514 Right CTATCTGCGTCTATCATCCAGCAC
6500 Right GATTCCGTCATATGGCTTGGATC
6515 Left AATTATAAGCAACAGAGGTGAAAAC
6501 Left GCAGGAGTCATGGGAGAAAAC 6516 Right CCGTAGCTCCAGACATCACT
6502 Right CTATCATCCAGCACTTGACCATG
6517 Left AGGTGAAAACAGCTGCAAGG
6503 Left CCAAGGGAGTTTGTGGAGAAC 6518 Right GTCATCAACTCCCAAACAGTCAC

6504 Right AAAGGTCATCAACTCCCAAACAG
6519 Left GGGAGTTTGTGGAGAACTCTG
6520 Right CATCCAGCACTTGACCATGATC
6521 Left CCATCCAAACTGCACCTACG
Sea 6522 Right TGATCAACTCACGGAACTTTGG
ID' Primer Sequence 6505 Left GAGAAAACAACACCCTGGTCTGGAA 6523 Left AATGGGCCTAAGATCCCGTC
6506 Right CGACTATCTGCGTCTATCATCCAG 6524 Right CGTCTATCATCCAGCACTTGAC
6507 Left AACGAATGGGCCTAAGATC CC
6508 Right CATTTCTATCAATGCAAGCCACG
Table 13. BRAF cDNA Capture Primer List for NGS Panel BRAF Regionl 75-125 bases Sea 6528 Right cactttc ccttgtagactgttc c ID' Primer Sequence 6525 Left gacaggaatcgaatgaaaacacttg 6529 Left ggaatcgaatgaaaacac 6526 Right Mc ccttgtagactgttc caaat 6530 Right ccactacccttgtagactgt Sea ID' Primer Sequence 6531 Left ggaatcgaatgaaaa 6527 Left ggaatcgaatgaaaacacttggtag 6532 Right ccactacccttgtagac BRAF Regionl 126-175 bases Sea 540 Right cattttcactgc cacatcac cat ID' Primer Sequence 6533 Left gaagacaggaatcgaatgaaaacac 6541 Left agacaggaatcgaatgaaaacacttg 6534 Right gctgtcacattcaacatiticactg 6542 Rjght Mee cttgtagactgttccaaat 6535 Left aggaatcgaatgaaaacacttggta 6543 Left ctcagaagacaggaatcgaatgaaa 6536 Right cacattcaacattttcactgccac 6544 Rjght catcaccatgccactttccc 6537 Left tatcatcctcagaagacaggaatc ¨ 545 Left aatc aat aaaacacttggtagac 6538 Right caacattttcactgccacatcac 6546 Right gctgtcacattcaacattttca __________________________________________ Seq.
Primer Sequence 6539 Left aagtcatcttcatcctcagaagaca ID

6547 Left aaatctccaggacctcagcgagaaa 6554 Right atcaccatgccacMccatgtag 6548 Right actttc cchgtagactgttc ca 6555 Left ctccaggacctcagcgag 6549 Left atcctcagaagacaggaatcgaa 6556 Right tgccacMccatgtagact 6550 Right atcaccatgccacMccatg 6557 Left catcttcatcctcagaagacagga 6551 Left cagcgagaaaggaagtcatct 6558 Right catcaccatgccacttt 6552 Right gccacMccatgtagactgtt 6559 Left ctcagcgagaaaggaagtca 6553 Left aagtcatchcatcctcagaag 6560 Right catcaccatgccac BRAF Regionl 176-225 bases S ID- Seq. . Primer Sequence Primer Sequence ID
6561 Left aagtcatchcatcctcagaagaca 6577 Left aagtcatcttcatcctcagaag 6562 Right gctgtcacattcaacatiticactg 6578 Right ctgctgaggtgtaggtgct 6563 Left actaactaacgtgaaagccttacag 6579 Left tcagcgagaaaggaagtca 6564 Right attttcactgccacatcaccat 6580 Right gchgtaactgctgaggtgta 6565 Left ccttacagaaatctccaggacct 6581 Left gaaatctccaggacctcagcgag 6566 Right tcacattcaacatiticactgcca 6582 Right tgctgtcacattcaacattt 6567 Left aagccttacagaaatctccagga 6583 Left aagtcatcttcatcctcag 6568 Right caacattttcactgccacatcac 6584 Right ttttgaaggcttgtaactgctg 6569 Left ctcagcgagaaaggaagtcatct 6585 Left ctcactaactaacgtgaaagcctta 6570 Right chgtaactgctgaggtgtaggt 6586 Right catcaccatgcca 6571 Left ttacagaaatctccaggacctcagc 6587 Left aagtcatcttcatcct 6572 Right tgctgtcacattcaacatitica 6588 Right aggcttgtaactgctgaggt 6573 Left ccaggacctcagcgagaaa 6589 Left ctcagcgagaaaggaag 6574 Right ctgaggtgtaggtgctgtca 6590 Right tgctgtcacattcaaca 6575 Left ctaacgtgaaagccttacagaaat 6591 Left agccttacagaaatctcca 6576 Right caacattttcactgccacat 6592 Right gctgtcacattca BRAF Regionl 226-275 bases Sea Primer Sequence 6602 Right chgtaactgctgaggtgtaggt 6593 Left agtcatcttcatcctcagaagaca 6603 Left attacct ctcactaactaac t 6594 Right tgaagagtaggatattcacatgtcg 6601 Rjght caacatiticactgccacatcac 6595 Left ctcactaactaacgtgaaagcctta 6605 Left ct ctcactaactaacgtgaaag 6596 Right cacattcaacattttcactgccac 6606 Right tgctgtcacattcaacattttca 6597 Left actaactaacgtgaaagccttacag 6607 Left aacgtgaaagccttacagaaatctc 6598 Right tittgaaggchgtaactgctgag 6608 Right ggcttgtaactgctgaggtgta 6599 Left cctcattacctggctcactaacta 6609 Left cttaca aaatctcca acctca 6600 Right gctgtcacattcaacattttcactg 6610 Right taactgctgaggtgtaggtgct 6601 Left actaacgtgaaagccttacagaaat 6611 Left aagccttacagaaatctccagga 6612 Right tittgaaggchgtaactgct 6622 Right gattcctgagtactcctac Sea 6623 Left cccctgcctcattacctg ID-. Primer Sequence 6624 Right tgctgtcacattcaacattt 6613 Left cctcattacctggctcactaa 6614 Right cattttcactgccacatcaccat 6625 Left aa tcatchcatcctca 6626 Ri ht t aa a ta atattcacatg 6615 Left gaaatctccaggacctcagcgagaa 6616 Right ctgaggtgtaggtgctgtca 6627 Left ctcagcgagaaaggaagtca 6628 Right Mgaaggcttgtaact 6617 Left agtcatchcatcctcagaag 6618 Right gcccatgaagagtaggatattc 6629 Left cctcattacctggctcac 6630 Right tgctgtcacattcaaca 6619 Left ctcagcgagaaaggaagtcatct 6620 Right catgtcgtgttnectga 6631 Left caggtngtctgctacccc 6632 Ri ht catcaccat cca 6621 Left aaatctccaggacctcagcg BRAF Regionl 276-325 bases Sea Primer Sequence 6633 Left aagtcatchcatcctcagaagaca 6634 Right ggaatagcccatgaagagtaggata 6653 Left aagccttacagaaatctccagga 6654 Right ggaatagcccatgaagagtagg 6635 Left actaactaacgtgaaagccttacag 6636 Right tgaagagtaggatattcacatgtcg 6655 Left tattgatgacttgattagagacca __________________________________________ 6656 Right caacatiticactgccacatcac 6637 Left ctcattacctggctcactaactaac 6638 Right tittgaaggchgtaactgctgag 6657 Left ttgattagagaccaaggatttcgtg __________________________________________ 6658 Right tgctgtcacattcaacatttt 6639 Left aacgtgaaagccttacagaaatctc 6640 Right atagcccatgaagagtaggatattc 6659 Left gctcactaactaacgtgaaagcctt 6660 Right attgaaggcttgtaactgct 6641 Left ttgatgacttgattagagaccaagg 6642 Right attttcactgccacatcaccat 6661 Left cttacagaaatctccaggacctca 6662 Right tgaagagtaggatattcacatg Sea 6663 Left cccctgcctcattacctgg ID-. Primer Sequence 6664 Right gtaactgctgaggtgtaggtgctg 6643 Left gatttcgtggtgatggaggatc 6644 Right gctgtcacattcaacatiticactg 6665 Left aaatctccaggacctcagcgagaaa 6666 ht cagagtggctngtggaat 6645 Left attacctggctcactaactaacgtg 6646 Right gchgtaactgctgaggtgtag 6667 Left ctcagcgagaaaggaagtcatct 6668 Ri ht taacaatagccagttgtg 6647 Left caaggatttcgtggtgatggag 6648 Right tcacattcaacattacactgcca 6669 Left atgacttgattagagaccaaggatt 6670 ht aacattacactgccacat 6649 Left cctcattacctggctcactaact 6650 Right cttgtaactgctgaggtgtaggtg 6671 Left ctaacgtgaaagccttacagaaat 6672 Right catgtcgtgtiticctgag 6651 Left ctggctcactaactaacgtgaaag 6652 Right ttgaaggchgtaactgctgaggtg BRAF Regionl 326-375 bases Sea Seq.
Primer Sequence Primer Sequence ID
6673 Left ctcattacctggctcactaactaac 6693 Left tattgatgacttgattagagacca 6674 Right ggaatagcccatgaagagtaggata 6694 Right cacattcaacatiticactgccac 6675 Left ctcactaactaacgtgaaagcctta 6695 Left gatttcgtggtgatggaggatc 6676 Right tgaagagtaggatattcacatgtcg 6696 Right gaaggcttgtaactgctgaggtgta 6677 Left agtcatcttcatcctcagaagaca 6697 Left aaacacaatagaacctgtcaatatt 6678 Right gatatggagatggtgatacaagctg 6698 Right tgctgtcacattcaacattttca 6679 Left acttgattagagaccaaggatttcg 6699 Left caaggatttcgtggtgatggag 6680 Right tittgaaggcttgtaactgctgag 6700 Right taactgctgaggtgtaggtgct 6681 Left aacgtgaaagccttacagaaatctc 6701 Left attacctggctcactaactaacgtg 6682 Right gaatagcccatgaagagtaggatattc 6702 Right cagagtggctngtggaat 6683 Left tgatgacttgattagagaccaagga 6703 Left actaactaacgtgaaagccttacag 6684 Right cttgtaactgctgaggtgtaggt 6704 Right ggtaacaatagccagttgtg 6685 Left actaacgtgaaagccttacagaaat 6705 Left cttacagaaatctccaggacctca 6686 Right ggaatagcccatgaagagtagg 6706 Right agccctcacaccactgg 6687 Left tataaacacaatagaacctgtcaat 6707 Left c cgatcctcatcagctc cc 6688 Right gctgtcacattcaacatiticactg 6708 Right caacatiticactgccacatca 6689 Left ctcagcgagaaaggaagtcatct 6709 Left ctggctcactaactaacgtgaaag 6690 Right atggagatggtgatacaagctggag 6710 Right tgaagagtaggatattcacatg 6691 Left gcatataaacacaatagaacctgtc 6711 Left ccaggacctcagcgagaaa 6692 Right cattttcactgccacatcacca 6712 Right tgatatggagatggtgatacaag BRAF Regionl 376-425 bases Sea 6724 Right atagcccatgaagagtaggatattc ID.' Primer Sequence 6713 Left actaactaacgtgaaagccttacag 6725 Left . atcatc aaatcaatttgggcaac 6714 Right gatatggagatggtgatacaagctg 672Ri ht cacattcaacatiticactgccac Seq.
Primer Sequence ID 6727 Left ctcactaactaacgtgaaagcctta 6715 Left cttgattagagaccaaggatttcgt 6728 Right atggagatggtgatacaagctggag 6716 Right ggaatagcccatgaagagtaggata 6717 Left aagtcatcttcatcctcagaagaca 6729 Left gcatataaacacaatagaacctgtc 6718 Right gcagtctgtcgtgcaatatctataa 6730 Right ttttgaaggcttgtaactgctgag 6719 Left gaagatcatcgaaatcaatttgggc 6731 Left cttc cgaccagcagatgaagatc 6720 Right gctgtcacattcaacatiticactg 6732 Right caacatiticactgccacatcac 6721 Left ttgatgacttgattagagaccaagg 6733 Left ctcagcgagaaaggaagtcatct 6722 Right tgaagagtaggatattcacatgtcg 6734 Right gcagtctgtcgtgcaatatcta 6723 Left tgacttgattagagaccaaggattt 6735 Left cctcattacctggctcactaacta 6736 Right tgatatggagatggtgatacaag 6745 Left actaacgtgaaagccttacagaaat 6737 Left ttacctggctcactaactaacgt 6746 Right ctggagccctcacaccac 6738 Right ttggtctcaatgatatggagatg 6747 Left aatctccaggacctcagcgagaaag 6739 Left gatttcgtggtgatggaggatc 6748 Right ctgtcgtgcaatatctataagtttg 6740 Right ggaatagcccatgaagagtagg 6749 Left gagaccgatcctcatcagctc 6741 Left ataaacacaatagaacctgtcaata 6750 Right aacattttcactgccacatcac cat 6742 Right cttgtaactgctgaggtgtaggt 6751 Left gaaatctccaggacctcagcgaga 6743 Left agaaatctccaggacctcagc 6752 Right tgatcatctcaaatttggtctcaa 6744 Right cgtgcaatatctataagtttgatca BRAF Regionl 426-475 bases Sea ID' Primer Sequence 6773 Left aa tcatchcatcctca aa aca 6753 Left actaacgtgaaagccttacagaaat 6774 Right ggatgattgacttggcgtgtaag 6754 Right gcagtctgtcgtgcaatatctataa 6775 Left . atcatc aaatcaatttgggcaac 6755 Left ccttcaaaatccattccaattccac 6776 Right chgtaactgctgaggtgtaggt 6756 Right gctgtcacattcaacatiticactg 6777 Left ctcactaactaacgtgaaagcctta 6757 Left attgatgacttgattagagaccaagg 6778 Right gcagtctgtcgtgcaatatcta 6758 Right ggaatagcccatgaagagtaggata 6779 Left gtccgtctecttcaaaatccatt 6759 Left actaactaacgtgaaagccttacag 6780 Rjght caacatiticactgccacatcac 6760 Right agtctgtcgtgcaatatctataagtt 6781 Left attacctggctcactaactaacgtg 6761 Left gaagatcatcgaaatcaatttgggc 6782 Ri ht t atcatctcaaatttggtctcaa 6762 Right tittgaaggchgtaactgctga 6783 Left atattgatgacttgattagagacca 6763 Left tgcatataaacacaatagaacctgtc 6784 Rjght aatagcccatgaagagtaggatattc 6764 Right tgaagagtaggatattcacatgtcg 6785 Left ctcagcgagaaaggaagtcatc 6765 Left caaaatccattccaattccacagc 6786 Right gaggtctctgtggatgattgactt 6766 Right cacattcaacattttcactgccac 6787 Left ttaca aaatctcca acctca 6767 Left aacgtgaaagccttacagaaatctc 6788 Right cttggcgtgtaagtaatccatgc 6768 Right gtgtaagtaatccatgccctgtg Sea 6789 Left caaattctcaccagtccgtctc ID' Primer Sequence 6790 Right aacattttcactgccacatcac cat 6769 Left gatttcgtggtgatggaggatc 6770 Right gatatggagatggtgatacaagctg 6791 Left ctggctcactaactaacgtgaaa 6792 Right cgtgcaatatctataagtttgatcatct 6771 Left ctcattacctggctcactaactaac 6772 Right cgtgcaatatctataagtttgatca BRAF Regionl 476-525 bases Sea 6795 Left tcactaactaacgtgaaagccttac ID' Primer Sequence 6796 Right ttattactettgaggtctctgtgga 6793 Left cttgattagagaccaaggatttcgt 6794 Right gatatggagatggtgatacaagctg 6797 Left . aa atcatc =
aaatcaatttgggc 6798 Right ggaatagcccatgaagagtaggata 6815 Left caaattctcaccagtccgtctc 6799 Left aagtcatcttcatcctcagaagaca 6816 Right gctgtcacattcaacatiticactg 6800 Right actgtagctagaccaaaatcaccta 6817 Left ctggctcactaactaacgtgaaag 6801 Left gatcatcgaaatcaatttgggcaac 6818 Right gtgtaagtaatccatgccctgtg 6802 Right tgaagagtaggatattcacatgtcg 6819 Left cttacagaaatctccaggacctca 6803 Left ctcattacctggctcactaactaac 6820 Right ggatgattgacttggcgtgtaag 6804 Right ctettgaggtctctgtggatgatt 6821 Left gtccgtctccttcaaaatccattc 6805 Left actaacgtgaaagccttacagaaat 6822 Right cttgtaactgctgaggtgtaggt 6806 Right gtctctgtggatgattgacttgg 6823 Left aaattctcaccagtccgtctccttc 6807 Left cttcaaaatccattccaattccaca 6824 Right cacattcaacatiticactgccac 6808 Right tittgaaggcttgtaactgctgag 6825 Left cttc cgaccagcagatgaagatc 6809 Left ctaactaacgtgaaagccttacaga 6826 Right atagcccatgaagagtaggatattc 6810 Right gaggtctctgtggatgattgact 6827 Left ttacctggctcactaactaacgtga 6811 Left gatttcgtggtgatggaggatc 6828 Right cttggcgtgtaagtaatccatgc 6812 Right gcagtctgtcgtgcaatatctataa 6829 Left caaggatttcgtggtgatggag Sea 6830 Right ctgtcgtgcaatatctataagtttg ID-. Primer Sequence 6813 Left tgatgacttgattagagaccaagga 6831 Left . act .
ccctaacatctggatcat 6814 Right atggagatggtgatacaagctggag 6832 Right atiticactgccacatcaccat BRAF Region2 75-125 bases Sea ID-. Primer Sequence Seq.Primer Sequence ID
6833 Left aatcatccacagagacctcaagag 6837 Left tcaatcatccacagagacctcaa 6834 Right tgttcaaactgatgggac cc 6838 Right tgttcaaactgatggga 6835 Left tccacagagacctcaagagtaa 6839 Left caagtcaatcatccacagagacc 6836 Right agacaactgttcaaactga 6840 Right tgggacccactc BRAF Region2 126-175 bases Sea 6850 Right agacaactgttcaaactgat ID-. Primer Sequence 6841 Left aatcatccacagagacctcaagag 6851 Left cttacacgccaagtcaatcatcc 6842 Right attctgatgacttctggtgccat 6852 Right aactgttcaaactgatggg Sea ID-. Primer Sequence 6853 Left gcatggattacttacacgccaag 6843 Left caagtcaatcatccacagagacc 6854 Right cacaaaatggatccagaca 6844 Right aactgttcaaactgatgggacc 6855 Left gccaagtcaatcatccacagag 6845 Left catccacagagacctcaagagtaa 6856 Right tgccatccacaaaatg 6846 Right cacaaaatggatccagacaact 6857 Left ttacacgccaagtcaatcatccaca 6847 Left tcaatcatccacagagacctcaa 6858 Right agacaactgttcaaact 6848 Right attctgatgacttctggtgc 6859 Left ggcatggattacttacacgcc 6849 Left cacagggcatggattacttacac 6860 Right cacaaaatggatccag 6861 Left ttatagatattgcacgacagactgc 6865 Left gcacgacagactgcacag 6862 Right tgggacccactc 6866 Right agacaactgttcaa 6863 Left cttacacgccaagtcaatca 6867 Left acgccaagtcaa 6864 Right tgccatccacaaa 6868 Right ctgatgacttctgg BRAF Region2 176-225 bases Sea Seq.
Primer Sequence Primer Sequence ID
6869 Left ttatagatattgcacgacagactgc 6887 Left gcatggattacttacacgcc 6870 Right attctgatgacttctggtgccat 6888 Right ctgactgaaagctgtatg 6871 Left cttacacgccaagtcaatcatcc 6889 Left gcacgacagactgcacag 6872 Right tctgactgaaagctgtatggatttt 6890 Right ttnatchgcattctgat 6873 Left tacacgccaagtcaatcatccacag 6891 Left gagatgatcaaacttatagatattgc 6874 Right atgcatatacatctgactgaaagct 6892 Right agacaactgttcaaact 6875 Left aacttatagatattgcacgacagac 6893 Left tgagaccaaatttgagatgatc 6876 Right cacaaaatggatccagacaact 6894 Right cacaaaatggatccag 6877 Left catggattacttacacgccaag 6895 Left cacagggcatggattactt 6878 Right tctgactgaaagctgtatggat 6896 Right attctgatgacttctgg 6879 Left cacagggcatggattacttacac 6897 Left acacgccaagtcaa 6880 Right attctgatgacttctggtgc 6898 Right aatgcatatacatctgactg 6881 Left aaacttatagatattgcacgaca 6899 Left catctccatatcattgagacca 6882 Right cacaaaatggatccagaca 6900 Right agacaactgttcaa 6883 Left acacgccaagtcaatca 6901 Left gagatgatcaaacttatagatat 6884 Right aatgcatatacatctgactgaaa 6902 Right tgccatccacaaa 6885 Left tgatcaaacttatagatattgcacg 6903 Left cacagggcatggatta 6886 Right tgccatccacaaaatg 6904 Right ttnatchgcattct BRAF Region2 226-275 bases Sea 6913 Left cacagggcatggattacttacac ID.' Primer Sequence 6914 Right acagaacaattccaaatgcatataca 6905 Left ttatagatattgcacgacagactgc 6906 Right tccaaatgcatatacatctgactga 6915 Left gcatggattacttacacgccaa Sea 6916 Right acaattccaaatgcatatacatctgac ID.' Primer Sequence 6907 Left aacttatagatattgcacgacagac 6917 Left tgatcaaacttatagatattgcacg 6908 Right tctgactgaaagctgtatggatttt 6918 Right tctgactgaaagctgtatggat 6909 Left tcaaacttatagatattgcacgaca 6919 Left cagettgtatcaccatctccatatc 6910 Right atgcatatacatctgactgaaagct 6920 Right ttctgatgacttctggtgc 6911 Left atcaccatctccatatcattgagac 6921 Left gcacgacagactgcacag 6912 Right attctgatgacttctggtgccat 6922 Right acagaacaattccaaatgcatat 6923 Left cacagggcatggattactta 6932 Right cacaaaatggatccag 6924 Right tccagtcatcaattcatacaga 6933 Left ttgagaccaaatttgagatgatc 6925 Left gagatgatcaaacttatagatattgc 6934 Right attctgatgacttctgg 6926 Right tctgactgaaagctgtatg 6935 Left ctccagettgtatcaccatctc 6927 Left tgtatcaccatctccatatcattga 6936 Right tgccatccacaaa 6928 Right tgccatccacaaaatg 6937 Left cacagggcatggat 6929 Left cacagggcatggattac 6938 Right acagaacaattccaaatgca 6930 Right caattccaaatgcatatacatct 6939 Left gagatgatcaaacttatagatat 6931 Left c cagettgtatcac catctc cat 6940 Right Matchgcattctga BRAF Region2 276-325 bases Sea ID' Primer Sequence 6961 Left ttgagaccaaatttgagatgatc 6941 Left cagcttgtatcaccatctccatatc 6962 Ri ht aca aacaattccaaat catataca 6942 Right tctgactgaaagctgtatggatttt 6963 Left cacagggcatggattactta 6943 Left atcaccatctccatatcattgagac 6964 Right ggtc cctgttgttgatgtttgaa 6944 Right atgcatatacatctgactgaaagct 6965 Left tgatcaaacttatagatattgcacgac 6945 Left acttatagatattgcacgacagactg 6966 Right ctgtccagtcatcaattcata 6946 Right ctgtccagtcatcaattcatacaga 6967 Left ccacaactggctattgttacc 6947 Left cacagggcatggattacttacac 6968 Right attctgatgacttctggtgc 6948 Right aaaattatctggtc cctgttgttga 6969 Left tt a accaaatttgagatg 6949 Left caaacttatagatattgcacgacaga 6970 Right acaattccaaatgcatatacatctg 6950 Right ccaaatgcatatacatctgactgaa 6971 Left ctccagettgtatcaccatctcc 6951 Left gcatggattacttacacgccaa 6972 Rjght tctgactgaaagctgtatg 6952 Right attatctggtcc ctgttgttgatgt 6973 Left gagatgatcaaacttatagat 6953 Left cacaactggctattgttacccag 6974 Ri ht aca aacaattccaaat catat 6954 Right attctgatgacttctggtgccat 6975 Left cacagggcatggattac 6955 Left gchgtatcaccatctccatatcatt 6976 Rjght aaaattatctggtcc ctgttgt 6956 Right tctgactgaaagctgtatggat 6977 Left tate ctactatcatgggctattcc 6957 Left gcacgacagactgcacag 6978 Rjght cacaaaatggatccag 6958 Right cctgagttgatgtttgaataaggt Sea 6979 Left attccacaaagccacaactg ID' Primer Sequence 6980 Right tgccatccacaaaatg 6959 Left gagatgatcaaacttatagatattgc 6960 Right attccaaatgcatatacatctgact BRAF Region2 326-375 bases Sea 6983 Left atcaccatctccatatcattgagac ID' Primer Sequence 6984 Right ctgtccagtcatcaattcatacaga 6981 Left ttatagatattgcacgacagactgc 6982 Right aaaattatctggtc cctgttgttga 6985 Left ccagettgtatcaccatctccatat 6986 Right tccaaatgcatatacatctgactga 7003 Left agatgatcaaacttatagatattgcac 6987 Left cacaactggctattgttacccag 7004 Right ggtecctgttgagatgtttgaa 6988 Right tctgactgaaagctgtatggatttt 7005 Left gchgtatcaccatctccatatcatt 6989 Left tatcctactatcatgggctattcc 7006 Right ctgtccagtcatcaattcatac 6990 Right attctgatgacttctggtgccat 7007 Left ccacaactggctattgttacc 6991 Left aacttatagatattgcacgacagac 7008 Right acaattccaaatgcatatacatctgac 6992 Right tatctggtecctgagttgatgat 7009 Left tgaatatcctactatcatgggcta 6993 Left gcatggattacttacacgccaa 7010 Right attctgatgacttctggtgc 6994 Right titiggacagnactccgtacctta 7011 Left agatgatcaaacttatagatattg 6995 Left cacagggcatggattacttacac 7012 Right cctgttgagatgatgaataaggt 6996 Right ccgtaccttactgagatctggag 7013 Left gcacgacagactgcacag 6997 Left tcaaacttatagatattgcacgaca 7014 Right aggtatcctcgtcccaccata 6998 Right ggtatcctcgtcccaccataaaa 7015 Left ttgagaccaaatttgagatgatc 6999 Left ctccagcttgtatcaccatctcc 7016 Right aaaattatctggtcc ctgttgt 7000 Right acagaacaattccaaatgcatataca 7017 Left ccagtggtgtgagggctc Sea 7018 Right tctgactgaaagctgtatggat ID Primer Sequence 7001 Left gctattccacaaagccacaac 7019 Left cacagggcatggattactta 7002 Right aatgcatatacatctgactgaaagc 7020 Right caggtatcctcgtcccacc BRAF Region2 376-425 bases Sea 7036 Right ctitiggacagttactccgtacc ID Primer Sequence 7021 Left cagcttgtatcaccatctccatatc 7037 Left ctccagettgtatcaccatctcc 7022 Right aaaattatctggtc cctgttgttga 70Ri ht attatctggtc cctgttgttgatgt 7023 Left tgaatatcctactatcatgggcta 7039 Left acgacatgtgaatatcctactct 7024 Right tctgactgaaagctgtatggatttt 7040 Ri ht tccaaat catatacatctgactga 7025 Left tatcctactatcatgggctattcc 7041 Left tcagcagttacaagccttcaaaa 7026 Right ctgtccagtcatcaattcatacaga 7042 Right ttctgatgacttctggtgc cat 7027 Left cgacatgtgaatatcctactatca 7043 Left tgatcaaacttatagatattgcacg 7028 Right atgcatatacatctgactgaaagct 7044 Right ccgtaccttactgagatctggag 7029 Left acttatagatattgcacgacagact 7045 Left cacagggcatggattacttacac 7030 Right titiggacagnactccgtacctta 7046 Right ggatttggacagttactccg Sea ID Primer Sequence 7047 Left gchgtatcaccatctccatatcatt 7031 Left atcaccatctccatatcattgagac 7048 Right aaaattatctggtcc ctgttgt 7032 Right ggtecctgagttgatgtttgaa 7049 Left gagatgatcaaacttatagatattgc 7033 Left ccacaactggctattgttaccc 7050 Right ggtatcctcgtcccaccataaaa 7034 Right cctgagttgatgtttgaataaggt 7051 Left tgaatatcctactatcatggg 7035 Left tcaaacttatagatattgcacgaca 7052 Right acagaacaattccaaatgcatataca 7053 Left ttgagaccaaatttgagatgatc 7057 Left attccacaaagccacaactg 7054 Right aggtatcctcgtcccaccata 7058 Right attccaaatgcatatacatctgac 7055 Left acctacacctcagcagttacaag 7059 Left gcacgacagactgcacag 7056 Right attctgatgacttctggtgc 7060 Right cactctgccattaatctatcat BRAF Region2 426-475 bases Sea ID- Seq. . Primer Sequence Primer Sequence ID
7061 Left tatcctactatcatgggctattcc 7081 Left tgtatcaccatctccatatcattga 7062 Right aaaattatctggtc cctgttgttga 7082 Right ccgtaccttactgagatctggag 7063 Left tgaatatcctactatcatgggcta 7083 Left tgacagcacctacacctcag 7064 Right ctgtccagtcatcaattcatacaga 7084 Right tctgactgaaagctgtatggatitt 7065 Left agettgtatcaccatctccatatca 7085 Left cacaactggctattgttacccag 7066 Right titiggacagnactccgtacctta 7086 Right ggtatcctcgtcccaccataaaa 7067 Left cgacatgtgaatatcctactatca 7087 Left gcatggattacttacacgccaa 7068 Right attatctggtcc ctgttgttgatgt 7088 Right aatagaggcgagaatttggggaaag 7069 Left atcaccatctccatatcattgagac 7089 Left tacacctcagcagttacaagc ctt 7070 Right chttggacagttactccgtacc 7090 Right acagaacaattccaaatgcatataca 7071 Left ctcagcagttacaagccttcaaaa 7091 Left aacttatagatattgcacgacagac 7072 Right tccaaatgcatatacatctgactga 7092 Right cactctgccattaatctatcat 7073 Left acctacacctcagcagttacaag 7093 Left acgacatgtgaatatcctactct 7074 Right atgcatatacatctgactgaaagct 7094 Right ggtecctgttgagatgtttgaa 7075 Left tatagatattgcacgacagactgc 7095 Left taggagtactcaggaaaacacgac 7076 Right gagaatttggggaaagagtggtc 7096 Right acagaacaattccaaatgcatat 7077 Left gatgtggcagtgaaaatgttgaatg 7097 Left ctccagettgtatcaccatctcc 7078 Right attctgatgacttctggtgccat 7098 Right ggatttggacagttactccg 7079 Left cagggcatggattacttacacg 7099 Left tcaaacttatagatattgcacgaca 7080 Right agtggtctctcatctettncttn 7100 Right agtggtctctcatctctitict BRAF Region2 476-525 bases Sea 7108 Right agtggtctctcatctctitictitt ID-. Primer Sequence 7101 Left cgacatgtgaatatcctactatca 7109 Left ttatagatattgcacgacagactgc 7102 Right aaaattatctggtc cctgttgttga 7110 Right aatagaggcgagaatttggggaaag _ Sea ID-. Primer Sequence 7111 Left gatgtggcagtgaaaatgttgaatg 7103 Left cagtgaaaatgttgaatgtgacagc 7112 Right ccaaatgcatatacatctgactgaa 7104 Right ctgtccagtcatcaattcatacaga 7105 Left tatcctactatcatgggctattcc 7113 Left atcaccatctccatatcattgagac 7106 Right Mggacagttactccgtacctta 7114 Right gagaatttggggaaagagtggtc 7107 Left gchgtatcaccatctccatatcatt 7115 Left tgaatatcctactatcatgggcta 7116 Right ggtatcctcgtcccaccataaaa 7129 Left ccagcttgtatcaccatctccatat 7117 Left catggtgatgtggcagtgaaaat 7130 Right cactctgccattaatctcttcat 7118 Right tctgactgaaagctgtatggatttt 7131 Left tggaacagtctacaagggaaagt 7119 Left tcagcagttacaagccttcaaaa 7132 Right ttctgatgacttctggtgc cat 7120 Right attatctggtcc ctgttgttgatgt 7133 Left aacagtctacaagggaaagtggc 7121 Left gtctacaagggaaagtggcatg 7134 Right attccaaatgcatatacatctgact 7122 Right atgcatatacatctgactgaaagct 7135 Left ctacacctcagcagttacaagcc 7123 Left aacttatagatattgcacgacagac 7136 Right ggtc cctgttgttgatgtttgaa 7124 Right aatagaggcgagaatttgggga 7137 Left tgacagcacctacacctcag 7125 Left ccacaactggctattgttaccc 7138 Right acagaacaattccaaatgcatataca 7126 Right cattggacagttactccgtacc 7139 Left cacagggcatggattacttacac 7127 Left acctacacctcagcagttaca 7140 Right aaggagggttctgatgcactg 7128 Right cctgagttgatgtttgaataaggt BRAF Region2 50-1250 bases Sea ID' Primer Sequence 7161 Left tgacttgattagagaccaaggattt 7141 Left taaatttcaccagcgttgtagtaca 7162 Right gagaatttggggaaagagtggtc 7142 Right aaaattatctggtc cctgttgttga 7163 Left aattat accaacttgatttgctgt 7143 Left catgtggttataaatttcaccagcg 7164 Right ggtc cctgttgttgatgtttgaa 7144 Right ctgtccagtcatcaattcatacaga 7165 Left cttgatttgctgthgtctccaag 7145 Left cttgattagagaccaaggatttcgt 7166 Right ggtatcctcgtcccaccataaaa 7146 Right titiggacagttactccgtacctta 7167 Left cctcattacctggctcactaactaa 7147 Left tatgaccaacttgatttgctgtttg 7168 Rjght aatagaggcgagaatttgggga 7148 Right cctgagttgatgtttgaataaggt 7169 Left caaaatccattccaattccacagc 7149 Left gaagatcatcgaaatcaatttgggc 7170 Ri ht catata actaaaatcctctgatgg 7150 Right agtggtctctcatctatttcttn 7171 Left cgttgtagtacagaagttccactg 7151 Left ccttcaaaatccattccaattccac 7172 Right tgagttgatgtttgaataaggtaac 7152 Right attatctggtcc ctgttgttgatgt 7173 Left gtctccttcaaaatccattccaatt 7153 Left tgatgacttgattagagaccaagga 7174 Right ggatttggacagttactccg 7154 Right cattggacagttactccgtacc 7175 Left cgtctccttcaaaatccattcca 7155 Left tggttataaatttcaccagcgttgt 7176 Right agcatatagactaaaatcctctgtt 7156 Right acagaacaattccaaatgcatataca Sea 7177 Left caacttgatttgctgtngtctcc ID-. Primer Sequence 7178 Right aggtatcctcgtcccaccata 7157 Left ttgtagtacagaagttccactgatg 7158 Right ccgtaccttactgagatctggag 7179 Left tgcatataaacacaatagaacctgt 7180 Right gcgagaatttggggaaagagtg 7159 Left gatcatcgaaatcaatttgggcaac 7160 Right aatagaggcgagaatttggggaaag Table 14. KRAS cDNA Capture Primer List for NGS Panel KRAS Regionl 75-125 bases Sea eq.
ID' Primer Sequence Primer Sequence ID
7181 Left GCCTGCTGAAAATGACTGAATATAA 7191 Left CTCAGCGGCTCCCAGG
7182 Right ATTCGTCCACAAAATGATTCTGAAT 7192 Right AAATGATTCTGAATTAGCTGTAT
7183 Left GAGAGAGGCCTGCTGAAAATG 7193 Left GTGCGGGAGAGAGGCC
7184 Right ATTGTTGGATCATATTCGTCCACAA 7194 Right AAATGATTCTGAATTAGCTG
7185 Left GGCCTGCTGAAAATGACTGAA 7195 Left GGGAGAGAGGCCTGCTG
7186 Right TCTATTGTTGGATCATATTCGTC CA 7196 Right TGTTGGATCATATTCGT
7187 Left GGAGAGAGGCCTGCTGAAA
7188 Right TGGATCATATTCGTCCACAAAATGA
7189 Left GAGAGGCCTGCTGAAAATGACT
7190 Right ATCATATTCGTCCACAAAATGATTC
KRAS Regionl 126-175 bases Sea ID' Primer Sequence 213 Left GCACTGAAGGCGGCG
Left GCCTGCTGAAAATGACTGAATATAA 7214 Right ATTCGTCCACAAAATGATTCTGAAT

7198 Right GTTTCTCCATCAATTACTACTTGCT 7215 Left GGGAGAGAGGCCTGCTG
___________________________________________ 216 Right ATCCTCTATTGTTGGATCATATT
7199 Left GAGAGAGGCCTGCTGAAAATG
7200 Right TATTGTTGGATCATATTCGTCCACA 7217 Left GGACTGGGAGCGAGCG
___________________________________________ 218 Right AAATGATTCTGAATTAGCTGTAT
7201 Left GAGAGGCCTGCTGAAAATGACTGAA _______ 7202 Right TCCTCTATTGTTGGATCATATTCGT 7219 Left GCTCCCAGGTGCGGGA
___________________________________________ 220 Right AGGAATCCTCTATTGTTGGA
7203 Left GGAGAGAGGCCTGCTGAAA
7204 Right TCCATCAATTACTACTTGCTTCCTG 7221 Left AGCGGCTCCCAGGTGC
___________________________________________ 222 Right ATCCTCTATTGTTGGATCAT
7205 Left GAGAGAGGCCTGCTGAAAATGACT
Right TATTGTTGGATCATATTCGTCCACAA 7223 Left CTGAAAATGACTGAATAT
7206 AA 7224 Right GAATATCCAAGAGACAGGTTTCT

7207 Left CTCAGCGGCTCCCAGG
7208 Right TGGATCATATTCGTCCACAAAATGA
7209 Left GTGCGGGAGAGAGGCC
7210 Right TCAATTACTACTTGCTTCCTGTAGG 225 Left CCAGAGGCTCAGCGG
7226 Right AAATGATTCTGAATTAGCTG
Seq. Primer Sequence ID 7227 Left TCAGCGGCTCCC
7211 Left CCAGGTGCGGGAGAGAG 7228 Right AGGAATCCTCTATTGTT
7212 Right ATCATATTCGTCCACAAAATGATTC
KRAS Regionl 176-225 bases S eq. Primer Sequence ID

7229 Left ATTTCGGACTGGGAGCGAG ID
7230 Right ATTGTTGGATCATATTCGTCCACAA 7247 Left AGCGGCTCCCAGGTGC
7248 Right GCTGTGTCGAGAATATCCAAGAG
7231 Left CTCGGCCAGTACTCCCG
7232 Right TGGATCATATTCGTCCACAAAATGA 7249 Left TCCCAGGTGCGGGAGAG
7250 Right CTCTTGACCTGCTGTGTCGA
7233 Left CAGGTGCGGGAGAGAGG
7234 Right TCGAGAATATCCAAGAGACAGGTTT 7251 Left CCAGAGGCTCAGCGG
7252 Right CCTGCTGTGTCGAGAATATCCAA
7235 Left CTCAGCGGCTCCCAGG
7236 Right TCCATCAATTACTACTTGCTTCCTG 7253 Left GCTCGGCCAGTACTC
7254 Right TCATATTCGTCCACAAAATGATTCT
7237 Left GCCAGTACTCCCGGCC
7238 Right ATTCGTCCACAAAATGATTCTGAAT 7255 Left CC CCGCCATTTCG
7256 Right TCCTCTATTGTTGGATCATATTCGT
7239 Left TGCGGGAGAGAGGCCT
7240 Right TGTGTCGAGAATATCCAAGAGACAG 7257 Left TCAGCGGCTCCC
7258 Right TTGACCTGCTGTGTCGAGAATATC
7241 Left GCACTGAAGGCGGCG
7242 Right TCAATTACTACTTGCTTCCTGTAGG 7259 Left GGCACTGAAGGCG
7260 Right ATCCTCTATTGTTGGATCATATT
7243 Left GCTCCCAGGTGCGGGA
7244 Right GTTTCTCCATCAATTACTACTTGCT 7261 Left CATTTCGGACTGGG
7262 Right AGGAATCCTCTATTGTTGGA
7245 Left CATTTCGGACTGGGAGC 7229 7246 Right TCTATTGTTGGATCATATTCGTCCA 7230 Left CCAGAGGCTCAG
Right GTTTCTCCATCAATTACTACTT
S eq. Primer Sequence KRAS Region2 75-125 bases Seq. 7242 Right TCCCCAGTCCTCATGTA
ID Primer Sequence 7231 Left CTTGGATATTCTCGACACAGCAG 7243 Left CTACAGGAAGCAAGTAGTAATTGA
7232 Right TTATGGCAAATACACAAAGAAAGCC 7244 Right TCCCCAGTCCTCAT
7233 Left AACCTGTCTCTTGGATATTCTCGAC 7245 Left AGAAACCTGTCTCTTGGATATT
7234 Right AAATACACAAAGAAAGCCCTCCC 7246 Right GATTTAGTATTATTTATGGC
7235 Left GGAAGCAAGTAGTAATTGATGGAGA 7 247 Left AGAAACCTGTCTCTTGGA
7236 Right AATACACAAAGAAAGCCCTCCCCAG 7248 Right GGCAAATACACAAAGAAA
7237 Left AGAAACCTGTCTCTTGGATATTCTC 7249 Left AGAAACCTGTCTCTT
7238 Right TCCCCAGTCCTCATGTACTG 7250 Right TTTATGGCAAATACACAAAG
7239 Left TGTCTCTTGGATATTCTCGACACAG 7251 Left CAAGTAGTAATTGATGG
7240 Right AAAGAAAGCCCTCCCCAGTCC 7252 Right ATGGCAAATACACA
Sea ID' Primer Sequence 7241 Left AGCAAGTAGTAATTGATGGAGAAAC
KRAS Region2 126-175 bases S eq. Primer Sequence ID

7253 Left TTTGTGGACGAATATGATCCAACAA ID
7254 Right TTATGGCAAATACACAAAGAAAGCC 7273 Left CAGGAAGCAAGTAGTAATTGATGGA
7274 Right GATTTAGTATTATTTATGGC
Left TGGACGAATATGATCCAACAATAGA
_______________________________________________________ 7255 G 7275 Left AGAAACCTGTCTCTTGGATATTCT
_ 7256 Right AAATACACAAAGAAAGCCCTCCC 7276 Right AATTTGTTCTCTATAATGGT
7257 Left CTTGGATATTCTCGACACAGCAG 7277 Left CTACAGGAAGCAAGTAGTAATTG
7258 Right AGGTACATCTTCAGAGTCCTTAAC 7278 Right TTTATGGCAAATACACAAAG
7259 Left TTCAGAATCATTTTGTGGACGAATA 7279 Left AGAAACCTGTCTCTTGGATAT
7260 Right AATACACAAAGAAAGCCCTCCCCAG 7280 Right GTCCTTAACTCTTTTAATTTG
7261 Left CCTTGACGATACAGCTAATTCAGAA 7281 Left ACAGCTAATTCAGAATCATTTTGTGG
7262 Right TCCCCAGTCCTCATGTACTG 7282 Right TCCCCAGTCCTCAT
7263 Left CATTTTGTGGACGAATATGATCCAA 7283 Left AGAAACCTGTCTCTTGG
7264 Right GAAAGCCCTCCCCAGTCC 7284 Right ACTCTTTTAATTTGTTCTCT
7265 Left TGTCTCTTGGATATTCTCGACACAG 7285 Left CTACAGGAAGCAAGTAGTAA
7266 Right GTCCTTAACTCTTTTAATTTGTTC 7286 Right TATAATGGTGAATATCTTC
7267 Left AACCTGTCTCTTGGATATTCTCGA 7287 Left TCCAACAATAGAGGATTCC
7268 Right TAATTTGTTCTCTATAATGGTGAA 7288 Right TTTATGGCAAATACACA
7269 Left GCAAGTAGTAATTGATGGAGAAAC 7289 Left AGAAACCTGTCTCT
7270 Right TTTATGGCAAATACACAAAGAAA 7290 Right GTCCTTAACTCTTTTAAT
7271 Left TGACGATACAGCTAATTCAGAATCA
7272 Right TCCCCAGTCCTCATGTA
S eq. Primer Sequence KRAS Region2 176-225 bases S ea ID'' Primer Sequence 7301 Left TACAGGAAGCAAGTAGTAA
7291 Left CCTTGACGATACAGCTAATTCAGAA 7302 Right CCATAGGTACATCTTCAGAGTCCTT
7292 Right TTATGGCAAATACACAAAGAAAGCC _ 7303 Left GCTAATTCAGAATCATTTTGTGGACG
7293 Left TTCAGAATCATTTTGTGGACGAATA 7304 Right TTTATGGCAAATACACAAAG
7294 Right GCAAATACACAAAGAAAGCCCTC
7305 Left GCAAGTAGTAATTGATGGAGAA
7295 Left CAGGAAGCAAGTAGTAATTGATGGA 7306 Right GTACATCTTCAGAGTCCTTAAC
7296 Right CCATAGGTACATCTTCAGAGTC
7307 Left GTGGACGAATATGATCCAACAATAG
_ 7297 Left TGACGATACAGCTAATTCAGAATCA 7308 Right AACTCTTTTAATTTGTTCTC
7298 Right TTTATGGCAAATACACAAAGAAA
7309 Left CTACAGGAAGCAAGTAGTAATTGA
7310 Right CCATAGGTACATCTTCAGA
S ea ID'' Primer Sequence 7311 Left TCATTTTGTGGACGAATATGATCCA
Left GGACGAATATGATCCAACAATAGAG 7312 Right GATTTAGTATTATTTATGGC

7300 Right TTTAATTTGTTCTCTATAATGGTG 7313 Left ACAGCTAATTCAGAATCATTTTGTGG

7314 Right TTTATGGCAAATACACA
7321 Left TACAGGAAGCAAGTAG
7315 Left TCCAACAATAGAGGATTCC 7322 Right GTCCTTAACTCTTTTAATTT
7316 Right GTCCTTAACTCTTTTAATTTGTT
7323 Left GAATATGATCCAACAA
7317 Left GAATCATTTTGTGGACGAATATGA 7324 Right GTCCTTAACTCTTTTAA
7318 Right ATAATGGTGAATATCTTC
7325 Left CTACAGGAAGCAAG
7319 Left AGAAACCTGTCTCTTG 7326 Right CCATAGGTACATCTTC
7320 Right CTAGAAGGCAAATCACATTTATTT
KRAS Region3 176-225 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
7327 Left gactgaatataaacttgtggtagt 7331 Left tgactgaatataaacttgt 7328 Right tc cc cagtcctcatgtactg 7332 Right cc ccagtcctcat 7329 Left gactgaatataaacttgtggt 7330 Right tc cc cagtcctcatgta KRAS Region3 226-275 bases Seq. Seq.
Primer Sequence Primer Sequence ID ID
7333 Left atgactgaatataaacttgtggtag 7343 Left GGGAGAGAGGCCTGCTG
7334 Right ttatggcaaatacacaaagaaagcc 7344 Right caaagaaagccctccccagtcct 7335 Left GCCTGCTGAAAatgactgaatataaa 7345 Left GGCCTGCTGAAAatgactgaata 7336 Right atacacaaagaaagcc ctcc cc 7346 Right cc ccagtcctcatgta 7337 Left GGAGAGAGGCCTGCTGAAAat 7347 Left TGCGGGAGAGAGGCCT
7338 Right gcaaatacacaaagaaagccctc 7348 Right tatggcaaatacacaaagaaa 7339 Left CCTGCTGAAAatgactgaatataaactt 7349 Left Aatgactgaatataaacttgtgg 7340 Right tc cc cagtcctcatgtactg 7350 Right tc cccagtcctcat 7341 Left GAGAGGCCTGCTGAAAatgact 7351 Left GGTGCGGGAGAGAGG
7342 Right atacacaaagaaagccctccccagt 7352 Right ggcaaatacacaaag KRAS Region3 276-325 bases Seq. . 7361 Left ATTTCGGACTGGGAGCGAG
Primer Sequence ID 7362 Right tccccagtcctcatgtactg 7353 Left CTCAGCGGCTCCCAGG
7354 Right ttatggcaaatacacaaagaaagcc 7363 Left GCACTGAAGGCGGCG
7364 Ri 7355 Left tgactgaatataaacttgtggtagt 7356 Right gtecttaactctittaatttgttc 7365 Left TGCGGGAGAGAGGCCT
7366 Ri ht Mat caaatacacaaagaaa 7357 Left TCCCAGGTGCGGGAGA
7358 Right aaatacacaaagaaagcc ctc cc _7367 Left GGCCTGCTGAAAatgactgaatata 7368 ht actatttaatttgttctct Seq. .
Primer Sequence ID 7369 Left AGCGGCTCCCAGGTGC
7359 Left GGCCTGCTGAAAatgactgaat 7370 Right cacaaagaaagccctccccagtcc 7360 Right tttaatttgttctctataatggtg __________________________________________ 7371 Left CCTGCTGAAAatgactgaatataaact 7372 Right gatttagtattatttatggc 7381 Left CCAGGTGCGGGAGAGAG
7373 Left tgactgaatataaacttgtggt 7382 Right Matggcaaatacaca 7374 Right acatcttcagagtecttaa 7383 Left CTGAAAatgactgaatataaacttgt 7375 Left GAGAGAGGCCTGCTGAAAatg 7384 Right gtecttaactctittaa 7376 Right tataatggtgaatatcttc 7385 Left GGCCTGCTGAAAatgact 7377 Left CATTTCGGACTGGGAGC 7386 Right ccttaactctittaatttg 7378 Right tc c ccagtcctcatgta 7387 Left GGCACTGAAGGCG
7379 Left CCAGAGGCTCAGCGG 7388 Right tccccagtcctcat 7380 Right Matggcaaatacacaaag KRAS Region3 326-375 bases Sea ID-Seq. . Primer Sequence Primer Sequence ID
7389 Left ATTTCGGACTGGGAGCGAG 7405 Left CATTTCGGACTGGGAGC
7390 Right ttatggcaaatacacaaagaaagcc 7406 Right aaagaaagccctccccagtcc 7391 Left CTCAGCGGCTCCCAGG 7407 Left AGCGGCTCCCAGGTGC
7392 Right ccataggtacatchcagagtectt 7408 Right gtecttaactchttaatttg 7393 Left CTCGGCCAGTACTCCCG 7409 Left GCTCGGCCAGTACTC
7394 Right aaatacacaaagaaagcc ctc cc 7410 Right tatggcaaatacacaaagaaa 7395 Left GTGCGGGAGAGAGGCC 7411 Left CCAGAGGCTCAGCGG
7396 Right taggtacatchcagagtecttaac 7412 Right actctittaatngttctct 7397 Left GCCAGTACTCCCGGCC 7413 Left CCCCGCCATTTCG
7398 Right aatacacaaagaaagccctccccag 7414 Right thatggcaaatacacaaag 7399 Left CCAGGTGCGGGAGAGAG 7415 Left TCAGCGGCTCCC
7400 Right ccataggtacatcttcagagtc 7416 Right ccataggtacatcttcaga 7401 Left GCTCCCAGGTGCGGGA 7417 Left GGCACTGAAGGCG
7402 Right gtecttaactctittaatttgac 7418 Right aatttgttctctataatggt 7403 Left GCACTGAAGGCGGCG
7404 Right taatttgactctataatggtgaa KRAS Region3 376-425 bases Sea ID-. Primer Sequence r425 Left CTCGGCCAGTACTCCCG
7419 Left CCAGGTGCGGGAGAGAG r426 Ri . ht Mat' . caaatacacaaa .
aaa . cc 7420 Right actgttctagaaggcaaatcacatt 7421 Left CTCAGCGGCTCCCAGG
7422 Right tctactgttctagaaggcaaatcac r 427 Left GTGCGGGAGAGAGGCC
7428 Right gtitigtgtctactgactagaagg Sea 7429 Left GCACTGAAGGCGGCG
ID.' Primer Sequence 7430 Right ccataggtacatchcagagtcat 7423 Left TTTCGGACTGGGAGCGAG
7424 Right aggtacatchcagagtccttaac 7431 Left TCCCAGGTGCGGGAGA

7432 Right tgactagaaggcaaatcacatttat 7445 Left GGCGGCGAAGGT
7446 Right atacacaaagaaagccctccccagt 7433 Left AGCGGCTCCCAGGTGC
7434 Right gcctgattgtgtctactgttc 7447 Left GCGAAGGTGGCG
7448 Right caaagaaagcc ctc cc cagtc ct 7435 Left GCCAGTACTCCCGGCC
7436 Right taatttgactctataatggtgaa 7449 Left TCAGCGGCTCCC
7450 Right tctactgactagaaggcaaat 7437 Left CATTTCGGACTGGGAGC 7419 7438 Right gtecttaactctittaatttgac 7420 Left CGGAGGCAGCAG
Right thatggcaaatacacaaagaaa 7439 Left CCAGAGGCTCAGCGG 7421 7440 Right ccataggtacatcttcagagtc 7422 Left CCCCGCCATTTCG
Right gtecttaactchttaatttg 7441 Left GCTCGGCCAGTACTC 7423 7442 Right ttatggcaaatacacaaagaaagccctc 7424 Left GGCACTGAAGGCG
Right ccataggtacatcttcaga 7443 Left AAGGTGGCGGCG 7425 7444 Right aaatacacaaagaaagcc ctc cc c 7426 Left CATTTCGGACTGGG
Right actctittaatngttctct KRAS Region3 426-475 bases Sea 7446 Right ttgctgatgatcaataaaaggaat ID Primer Sequence 7427 Left ATTTCGGACTGGGAGCGAG _7447 Left CATTTCGGACTGGGAGC
7428 Right actgttctagaaggcaaatcacatt 7448 Right tgactagaaggcaaatcacatttat 7429 Left GCTCGGCCAGTACTCCC 7449 Left GGCACTGAAGGCG
7430 Right ccataggtacatchcagagtectt 7450 Rjght ttchgctaagtectgagcctgat 7431 Left GTGCGGGAGAGAGGCC 7451 Left GCTCGGCCAGTACT
7432 Right gtchgtattgctgatgatcaat _7452 Right ccataggtacatcttcagagtc 7433 Left GCCAGTACTCCCGGCC 7453 Left AAGGTGGCGGCG
7434 Right taggtacatchcagagtecttaac 7454 Right gtecttaactchttaatngttc 7435 Left CTCAGCGGCTCCCAGG 7455 Left GGCGGCGAAGGT
7436 Right agcctgtitigtgtctactgttc 7456 Ri ht taattt tctctataatggtgaa 7437 Left GCACTGAAGGCGGCG 7457 Left TCAGCGGCTCCC
7438 Right tctactgttctagaaggcaaatcac 7458 Ri ht aattccataacttchgctaag 7439 Left CCAGGTGCGGGAGAGAG 7459 Left CATTTCGGACTGGG
7440 Right cttchgctaagtectgagcct 7460 Rjght tctactgactagaaggcaaat 7441 Left GCTCCCAGGTGCGGGA 7461 Left CCCCGCCATTTCG
7442 Right tgctgatgatcaataaaaggaattcc 7462 Ri ht ccata tacatcttcaga 7443 Left CCAGAGGCTCAGCGG 7463 Left GCGAAGGTGGCG
7444 Right gttngtgtctactgactagaagg 7464 Right gtecttaactchttaatttg Sea ID Primer Sequence 7465 Left CCAGAGGCTCAG
7445 Left AGCGGCTCCCAGGTGC 7466 Right agcctgattgtgtctactg KRAS Region3 476-525 bases Sea Seq.
Primer Sequence Primer Sequence ID
7467 Left GCTCGGCCAGTACTCCC 7487 Left CCAGAGGCTCAGCGG
7468 Right actgttctagaaggcaaatcacatt 7488 Right tgctgatgtttcaataaaaggaattcc 7469 Left CATTTCGGACTGGGAGCGA 7489 Left GCTCGGCCAGTACT
7470 Right gttngtgtctactgactagaagg 7490 Right tctactgactagaaggcaaatcac 7471 Left CAGGTGCGGGAGAGAGG 7491 Left CATTTCGGACTGGGAG
7472 Right atgtatagaaggcatcatcaacacc 7492 Right cacttgctaagtectgagcct 7473 Left CTCAGCGGCTCCCAGG 7493 Left GGCGGCGAAGGT
7474 Right gtchgtattgctgatgatcaat 7494 Right ccataggtacatchcagagtcat 7475 Left TCCCAGGTGCGGGAGA 7495 Left AAGGTGGCGGCG
7476 Right tatagaaggcatcatcaacaccctg 7496 Right tgactagaaggcaaatcacatttat 7477 Left TGCGGGAGAGAGGCCT 7497 Left GGCACTGAAGGCG
7478 Right atcatcaacaccctgtchgtatt 7498 Right tchgtattgctgatgatcaataaa 7479 Left GCCAGTACTCCCGGCC 7499 Left TCAGCGGCTCCC
7480 Right agcctgtitigtgtctactgttc 7500 Right ttgctgatgatcaataaaaggaat 7481 Left GCACTGAAGGCGGCG 7501 Left CGGAGGCAGCAG
7482 Right ctgtchgtattgctgatgatc 7502 Right taggtacatcttcagagtc cttaac 7483 Left GGACTGGGAGCGAGCG 7503 Left CCCCGCCATTTCG
7484 Right ttchgctaagtc ctgagc ctgttt 7504 Right tctactgactagaaggcaaat 7485 Left AGCGGCTCCCAGGTGC 7505 Left GCGAAGGTGGCG
7486 Right catcatcaacaccctgtchgtc 7506 Right gtitigtgtctactgactaga KRAS Region3 526-575 bases Sea 7516 Right tatagaaggcatcatcaacaccctg ID.' Primer Sequence 7507 Left ATTTCGGACTGGGAGCGAG 7517 Left CATTTCGGACTGGGAGC
7508 Right atgtatagaaggcatcatcaacacc 751Ri ht atcatcaacaccctgtchgtatt 7509 Left GCTCGGCCAGTACTCCC 7519 Left CTCAGCGGCTCCCAGG
7510 Right gtchgtattgctgatgatcaat _7520 Right attaccatcMgctcatctt 7511 Left GCCAGTACTCCCGGCC 7521 Left GCTCGGCCAGTACT
7512 Right ctgtchgtattgctgatgatc 7522 Right tgctgatgtttcaataaaaggaattcc 7513 Left GTGCGGGAGAGAGGCC
7514 Right tittaccatcMgctcatctitic 7523 Left AGCGGCTCCCAGGTGC
7524 Right tgctcatctntattatgat Sea ID.' Primer Sequence 7525 Left GGCGGCGAAGGT
7515 Left GCACTGAAGGCGGCG

7526 Right gttngtgtctactgactagaagg 7537 Left GCGAAGGTGGCG
7527 Left CCCCGCCATTTCG 7538 Right agcctgattgtgtctactgttc 7528 Right tatgtattgctgatgthcaataaa 7539 Left CCAGGTGCGGGAGAGAG
7529 Left AAGGTGGCGGCG 7540 Right Mac catcMgctcat 7530 Right ttgctgatgatcaataaaaggaat 7541 Left CCAGAGGCTCAGCGG
7531 Left GGCACTGAAGGCG 7542 Right gaatttctcgaactaatgtata 7532 Right catcatcaacaccctgtchgtc 7543 Left CATTTCGGACTGGG
7533 Left GCTCCCAGGTGCGGGA 7544 Right cctgtettgtattgctgatgt 7534 Right attatgattcgaatnctc 7545 Left TCAGCGGCTCCC
7535 Left CGGAGGCAGCAG 7546 Right ttgctcatcttnctttatg 7536 Right actgttctagaaggcaaatcacatt KRAS Region3 576-625 bases Sea ID-Seq. . Primer Sequence Primer Sequence ID
7547 Left GCTCGGCCAGTACTCCC 7565 Left CAGGTGCGGGAGAGAGG
7548 Right atgtatagaaggcatcatcaacacc 7566 Right ttacataattacacactttg 7549 Left CATTTCGGACTGGGAGCGA 7567 Left CCCCGCCATTTCG
7550 Right tittaccatcMgctcatctitic 7568 Right taatgtatagaaggcatcatcaac 7551 Left GCCAGTACTCCCGGCC 7569 Left CTCAGCGGCTCCCAGG
7552 Right tatagaaggcatcatcaacaccctg 7570 Right cacactagtattgac 7553 Left GGACTGGGAGCGAGCG 7571 Left CGGAGGCAGCAG
7554 Right tittaccatcMgctcatch 7572 Right cctgtettgtattgctgatgt 7555 Left GCACTGAAGGCGGCGG 7573 Left CATTTCGGACTGGGAG
7556 Right tgctcatctitictnatgat 7574 Right gaatttctcgaactaatgtata 7557 Left GCTCGGCCAGTACT 7575 Left GGCACTGAAGGCGG
7558 Right atcatcaacaccctgtchgtatt 7576 Right attaccatcMgctcat 7559 Left AAGGTGGCGGCG 7577 Left CTCCCAGGTGCGGGA
7560 Right gtchgtattgctgatgatcaat 7578 Right gtattgacttatt 7561 Left GGCGGCGAAGGT 7579 Left CATTTCGGACTGG
7562 Right ctgtchgtattgctgatgatc 7580 Right ctttatgtiticgaatttctc 7563 Left GCGAAGGTGGCG
7564 Right tgctgatgatcaataaaaggaattcc KRAS Region3 626-675 bases Sea ID
ID-. Primer Sequence 7583 Left GCCAGTACTCCCGGCC
7581 Left GCTCGGCCAGTACTCCC 74 Rjght attaccatcMgctcatctt 7582 Right tittaccatcMgctcatctitic Seq. Primer Sequence 7585 Left ATTTCGGACTGGGAGCGAG

7586 Right ttacataattacacactttg 7595 Left AAGGTGGCGGCG
7596 Right ctttatgtiticgaatttctc 7587 Left CGGAGGCAGCAG
7588 Right atgtatagaaggcatcatcaacacc 7597 Left CCCCGCCATTTCG
7598 Right ttttaccatctttgctcat 7589 Left GCTCGGCCAGTACT
7590 Right tgctcatctitictnatgat 7599 Left GCGAAGGTGGCG
7600 Right ttgctcatcttttctttatg 7591 Left CATTTCGGACTGGGAGC
7592 Right cacactngtattgac 7601 Left CATTTCGGACTGGG
7602 Right gtattgacttatt 7593 Left GGCGGCGAAGGT
7594 Right gaatttctcgaactaatgtata KRAS Region3 676-725 bases Sea Seq.
ID.' Primer Sequence ID Primer Sequence 7603 Left GCTCGGCCAGTACTCCC 7607 Left GCTCGGCCAGTACT
7604 Right ttacataattacacactttg 7608 Right cacactagtattgac 7605 Left CGGAGGCAGCAG 7609 Left AAGGTGGCGGCG
7606 Right tittaccatcMgctcatctitt 7610 Right gtattgacttatt Table 15: IRID Hybridization-based capture probes for NGS panel ID Probe Sequence CCATTTCCATCCTGCAGAAGAAGCCCCGCCACCAG

TGCTAACGATCTCTTTGATGATGGCTGTCATGTCTGGGAGCCTGTGGCTGAAGAAAAAG
GAGGAGAGAGATGGCAGAAGCTGC

GGGGCTGAGGTGGAAG

CTCCGGGCCTGTCTCTTCCACCTCAGCCCCTCAAGAATGGGG

TGGGAAGAAAGGAAATGCATTTCCTAATTTTATCCCTAGGAAGATGAGTGTACAACGG

TGTCCCGAGCCACGTGCAGAAGGTCCAGCATGGCCAGGGAGGAGGGCTGGCTCTGTGG
GGAGACAGAAGCGGGC

GCTCCTGTCCTTGGC

GCCCTCGCCCGGTGAGTGAGAACCAGTCTTTGCTGCAGTTGTTGTGCC

TTGCTGCAGAGCAGAGAGGGATGTAACCAAAATTAACTGAGCTGAGTCTGGGCAAATC
TTAAACTGGGAGGAACA

GCCCCTACACTGCACCCCTCTCCTCCCAGGACGGC

GAAAAACATCACCCTCATTCGGTGAGCGCCCTGCTGC

TGCAGCTCCATCTGCATGGCTTGCAGCTCCTGGTGCTTCCGGCGGTACACTGCAGGTGG
GTG

AATTCAAGAGTTTTTTTTTCTTATTCTGAGGTTATCTTTTTACCACAG

TGGTCTTCAAAAGGATATTGTGCAACT

GGTGTAATGATATGTGCATATTTATTACATCGGGGCAA

CCCATAGAAATCTAGGGCCTCTTGTGCCTTTAAAAATT

CTCTTTTTTTTCTGTCCACCAGGGAGTAACTA

TAGCTATAATAATACACATAGCGCCTCTGACTGGGAAT

CTTCCATGTATTGGAACAGTTTTCTTAACCATATCTAGAAGTTTACATAAAAATTTAGA
AAGAAATTT

AAGGCATTTCCTGTGAAATAATAC

AGGATCTGCATGGTTAAATACATACCAGTATT

ATCAAAGTAGAGTTCT

ATTTCTCCCAATGAAAGTAAAGTACAAACCTTTTTTAGCATCTTGTTCTGTTTGTGGAA

ATTTCATTTCTTTTTCTTTTCTTTTTTTTTTTTTTTAGGACAAAATGTTTCACTTTTGGGTA

TGTATTTA

ATATCTAGTACTTACTTTAACAAAAAATGATCTTGACAAAGCAAATAAAGACAA

GTATCGGTTGGCTT

AAGATTTACCTCTATTGTTGGATCA

GAATATG

TGAAAATGTGACTATATTAGAACATGTCACACATAAGGTTAATACA

GGTGAGGGCTGAGGTGACCCTTGTCTCTGTGTTCTTGTCCCCCCCAGCTTGTGGAGCCT
CTTACACC

CACTTTGATCTTTTTGAATTCAGTTTCCTTCAAGATCCTCAAGAGAGCTTGGTTGGGAGC
TTCTCCACTGG

CTCACAATTGCCAGTTAACGTCTTCCT

CCCTATGACAGAGAGAGAAG

TGGGGGTCCATGGCTCTGAACCTCAGGCCCACCTTTTCTCATGTCTGGCAGC

CCTTCTGGCCACCATGCGAAG

CGTAGGCTTCCTGGAGGGAGGGAGAGGCACGTCAGTGTGGC

GAACACAAAGACAATATTGGCTCCCAGTACCTGCTCAACTGGTGTGTGCAGATCGCAA
AGGTAATCAGGGAAGGGA

TCCCATGATGATCTGTCCCTCACAGCAGGGT

TCCAAGTAGTTCATGCCCTGAAACAGAGAAGACCCTGC

AATACCATGCAGAAGGAGGCAAAGTAAGGAGGTGGCTTTAGGTCAGCCAGCATTT

TTGACTTCTCTGGTGACAG

AAGTGGGGAGCACAAGTCAATACT

AAGGATGACGACTTTGAGAAGATCAGTGAGCTGGGGGCTGGCAATGGCGGTGTGGTGT

TCAAG

ACCTTTCTGGCCATGACCAGGCCAGAAGGCTTGTGGGAGACC

AAAAATTTAATCAGTGGAAAAATAGCCTCA

TTGTGGATGGTAAGAATT

GTAGTAAGTAAAGGAAAACAGTAGAT

AAAACCTTACTTTATTTGGATTTGAT

TGGTTCAGCAGTGTGGTAAAGT

TAAATCCTGCTTCTCGGGATACAGACCAATTGGCATGCTCTTCAATCACTGACATATCT
GGGAA

AGGTTGGCCTGGACCC

TTTGGGGGCAGAG

CTCTGCCCCCAAAGAAC

GGGAGGGTGGGGAGGAGGAGGAGGCTGTGA

CTGTAGGCTGCAGTTCTCAGCTC

GCGCCATGGCCCAGACCCCTGTGCAAAGGAGAAGACAAGAGG

AAGAGCTCTACCAATGTGAGTGACCATTATCACTCCT

AGTTTGCCTTCCAGAACATCCTCACATGTA

TATGCAATGGACCGACCGTGATCAGATTAGGGTTACCTGAGGATCGAATGAATTGAAA
TG

GTTTTAGAAAATGAATAGTTGTTAAACCTGAATT

AAATTCAGGTTTAACAACTATTCATTTT

ACTCTTGCTCTGCTAGCCTATCTAGCTAAAGAGCTTCTCTG

GCAGCCTGTACCCAGTGGTGCCGAGCCTCTGGCG

GTGGATCTGTGGGTGGGGGTGGTGTGAGGCTTGGCACCG

GCCACCCATCTCCTCAGCTGAGATGAC

TGACCGGGAAGGCCTGGGCGGGCCTCCGGAAGG

CCTCCCCAGCTGCCTTCCAGGGAGGGAGGCCAGCTGG

CTCCTTTACCCCTCCTTCCTAGAGAGTTAGA

TAAAGACATTGAAGTTACTC

GGAGGTTACAGAAACACCCTTATACATGTAGTGGGGCCACGACGCCCGTCTGTGCAGC
TTGGCCAG

CAGGTCAGCGGGCTACCACTGGGCC

TGAATATGATCCCACCATAGAGGTGAGG

CCAGTAATCAGGGTTAATTGGCGAGCCACATCTACAGTACTTTAAAGCTTTCTATAATC
A

TGTATTTGCCAT

CACTTGTTTTCTGTAAGAATCCTGGGGGTGT

TGCCCCCTTACCCTCCA

TGCTAATATATAATCACAAACAAAAAGTATCCAATATCACCCTACATAAAAGAAAACC
C

TAGGTGGGTTTAAATTGAATATAATAAGCTGACATTAAGGAGTAATTATAGTTTTTATT
TTTTGAG

CGAGAATATCCAAGAGACAGGT

GCACTGTAATAATCCAGACTGTGTTTCTCCCTTCTCAGGATTCCTACAGGAAGCAAGTA
GTAATTGATGGAGAAA

GTGGGCAAAGAGGGCTCCAGCCAACCCCCCAAAT

GTGGCGCGTAAGTATCCCCTTGGCCTCTCGGGATTCAGATTTGGGGGGTTGGCTGGAGC
CC

TACTCCCCTACAGACGTGCGGGTGGTGAGAGCCACGCACACTCTACCCGTCAGACCCTC
GCCAGGCAGCCAGG

AAAATTAGAACAGTAGATGCTTAGTTTA

GCAGTCAAAGAGAGTTAGAAAAGCAT

GTATCAGAATCTCTGTGCCACAATCCAAATTAAGTGACAAGGAGGAATCTG

TTCTGATCTGGTTGAA

TTCATTTTTAGAAGTTA

ATGACTGTGTTCTTAAGGTA

TTTAACAAGCTCTTT

TAATTCACTGCCCAGATCTTAAAACAGAGAGAAAGA

CCAGAAGGTATATTTCAGTTTATTGTTCTGAGAAATACCTATACAT

AAGGATGACGACTTTGAGAAGATCAGTGAGCTGGGGGCTGGCAATGGCGGTGTGGTGT
CCAAA

GTATCTAGTACTTACTTTAACAAAAAATGATCTTGACAAAGCAAATAAAGACAA

TGTATTTACCCAAAAGTGAAACATTTTGTCCTTTTTTAGCATCTTGTTCTGTTTGTGGAA

ATCAAAGTAGAGTTCTTCCACAAACAGAACAAGATGCTAAAAAAGGACAAAATGTTTC
ACTTTTGGGTA

GCTAAAGGTGAAGATGTATTCCTCCAATTCAGGACCCACACGATGGGAGGACAAGTTC
ATGTATTTTGAGTTCCCTCAGCCGTTACCTGTGTGTGGTGATATCAAAGTAGAGTTCT

TAGCTATAGTAATACACATAGCGCCTCTGACTGGGAATAGTTACTCCCTTTTTGTCTCTG
GTCCTTACTTCCCCATAGAAATCTAGGGCCTCTTGTGCCTTTAAAAATT

GGTATAATGATTTATGCATATTTATTACATCGGGGCAA

CATTTCCATCCTGCAGAGCCCCGCCACCAG

CAATATGATCCAACAATAGAGAATTCCTACAGGAAGCAAGTAGTAATTGATGGAGAAA

ACCTGTTGTGTCAAGAATATCCAAGAGACAGGT

References 1. Bang Y, Kwak EL, Shaw AT, Camidge DR, Iafrate AJ, Maki RG, Solomon BJ, Ou R, Salgia R, Clark JW. Activity of the oral ALK inhibitor PF-02341066 in ALK-positive patients with non-small cell lung cancer (NSCLC). J Clin Oncol 28:
18s (suppl; abstr 3), 2010.
2. Butrynski JE, D'Adamo DR, Hornick JL, Dal CM P, Antonescu CR, Jhanwar SC, Ladanyi M, Capelletti M, Rodig SJ, Ramaiya N, Kwak EL, Clark JW, Wilner KD, Christensen JG, Janne PA, Maki RG, Demetri GD, Shapiro GI. Crizotinib in ALK-rearranged inflammatory myofibroblastic tumor. N Engl J Med. 2010 Oct 28;363(18):1727-33.
3. Camidge DR, Doebele RC.Treating ALK-positive lung cancer--early successes and future challenges. Nat Rev Clin Oncol. 2012 Apr 3;9(5):268-77.
4. Cheng M, Ott GR. Anaplastic lymphoma kinase as a therapeutic target in anaplastic large cell lymphoma, non-small cell lung cancer and neuroblastoma. Anticancer Agents Med Chem 10: 236-249, 2010.
5. Choi YL, Soda M, Yamashita Y, Ueno T, Takashima J, Nakajima T, Yatabe Y, Takeuchi K, Hamada T, Haruta H, Ishikawa Y, Kimura H, Mitsudomi T, Tanio Y, Mano H; ALK Lung Cancer Study Group. EML4-ALK mutations in lung cancer that confer resistance to ALK inhibitors. N Engl J Med. 2010 Oct 28;363(18):1734-9.
6. Choi YL, Soda M, Yamashita Y, Ueno T, Takashima J, Nakajima T, Yatabe Y, Takeuchi K, Hamada T, Haruta H, Ishikawa Y, Kimura H, Mitsudomi T, Tanio Y, Mano H. ALK Lung Cancer Study Group. EML4-ALK mutations in lung cancer that confer resistance to ALK inhibitors. N Engl J Med. 2010 Oct 28;363(18):1734-9.
7. Christensen JG, Zou HY, Arango ME, Li Q, Lee JH, McDonnell SR, Yamazaki S, Alton GR, Mroczkowski B, Los G. Cytoreductive antitumor activity of PF-2341066, a novel inhibitor of anaplastic lymphoma kinase and c-Met, in experimental models of anaplastic large-cell lymphoma. Mol Cancer Ther 6 (12, Pt. 1): 3314-3322, 2007.
8. Cui JJ et al. Structure based drug design for the discovery of clinical candidate PF-2341066 as potent and highly selective c-Met inhibitor. Abstracts of Papers, 235th ACS National Meeting, New Orleans, LA, United States, April 6-10, 2008, 2008:
p.
MEDI-177.
9. Doebele RC, Pilling AB, Aisner DL, Kutateladze TG, Le AT, Weickhardt AJ, Kondo KL, Linderman DJ, Heasley LE, Franklin WA, Varella-Garcia M, Camidge DR. Mechanisms of resistance to crizotinib in patients with ALK gene rearranged non-small cell lung cancer. Clin Cancer Res. 2012 Mar 1;18(5):1472-82.
10. Doebele RC, Pilling AB, Aisner DL, Kutateladze TG, Le AT, Weickhardt AJ, Kondo KL, Linderman DJ, Heasley LE, Franklin WA, Varella-Garcia M, Camidge DR. Mechanisms of resistance to crizotinib in patients with ALK gene rearranged non-small cell lung cancer. Clin Cancer Res. 2012 Mar 1;18(5):1472-82. Epub Jan 10.
11. Engelman JA, Zejnullahu K, Mitsudomi T, Song Y, Hyland C, Park JO, Lindeman N, Gale CM, Zhao X, Christensen J, Kosaka T, Holmes AJ, Rogers AM, Cappuzzo F, Mok T, Lee C, Johnson BE, Cantley LC, Janne PA. Science. 2007 May 18;316(5827):1039-43. Epub 2007 Apr 26.
12. Hallberg B, Palmer RH. Crizotinib¨latest champion in the cancer wars? N
Engl J
Med 363: 1760-1762, 2010.
13. Koivunen JP, Mermel C, Zejnullahu K, Murphy C, Lifshits E, Holmes AJ, Choi HG, Kim J, Chiang D, Thomas R, Lee J, Richards WG, Sugarbaker DJ, Ducko C, Lindeman N, Marcoux JP, Engelman JA, Gray NS, Lee C, Meyerson M, Janne PA.
EML4-ALK fusion gene and efficacy of an ALK kinase inhibitor in lung cancer.
Clin Cancer Res 14: 4275-4283, 2008.
14. Loyly CM, Pao. Escaping ALK inhibition: mechanisms of and strategies to overcome resistance. W. Sci Transl Med. 2012 Feb 8;4(120):120ps2.

15. McDermott U, Iafrate AJ, Gray NS, Shioda T, Classon M, Maheswaran S, Zhou W, Choi HG, Smith SL, Dowell L, Ulkus LE, Kuhlmann G, Greninger P, Christensen JG, Haber DA, Settleman J. Genomic alterations of anaplastic lymphoma kinase may sensitize tumors to anaplastic lymphoma kinase inhibitors. Cancer Res 68:
3389-3395, 2008.

16. Sasaki T, Okuda K, Zheng W, Butrynski J, Capelletti M, Wang L, Gray NS, Wilner K, Christensen JG, Demetri G, Shapiro GI, Rodig SJ, Eck MJ, Janne PA. The neuroblastoma associated F1174L ALK mutation causes resistance to an ALK
kinase inhibitor in ALK translocated cancers. Cancer Res. 2010 Dec 15;70(24):10038-43.

17. Sasaki T, Rodig SJ, Chirieac LR, Janne PA. The biology and treatment of ALK non-small cell lung cancer. Eur J Cancer 46: 1773-1780, 2010.

18. Shaw AT, Yeap BY, Mino-Kenudson M, Digumarthy SR, Costa DB, Heist RS, Solomon B, Stubbs H, Admane S, McDermott U, Settleman J, Kobayashi S, Mark EJ, Rodig SJ, Chirieac LR, Kwak EL, Lynch TJ, Iafrate AJ. Clinical features and outcome of patients with non-small-cell lung cancer who harbor EML4-ALK. J
Clin Oncol 27: 4247-4253, 2009.

19. Wood AC, Laudenslager M, Haglund EA, Attiyeh EF, Pawel B, Courtright J, Plegaria J, Christensen JG, Mosse YP. Inhibition of ALK mutated neuroblastomas by the selective inhibitor PF-02341066. J Clin Oncol 27:15s, 2009 (suppl;
abstr 10008b).

20. Zou HY, Li Q, Lee JH, Arango ME, McDonnell SR, Yamazaki S, Koudrakova TB, Alton G, Cui JJ, Kung PP, Nambu MD, Los G, Bender SL, Mroczkowski B, Christensen JG. An orally available small-molecule inhibitor of c-Met, PF-2341066, exhibits cytoreductiye antitumor efficacy through antiproliferatiye and antiangiogenic mechanisms. Cancer Res 67: 4408-4417, 2007.
SEQUENCES
anaplastic lymphoma kinase (ALK) SEQ ID NO: 7714 1 mgaigllwl1p111staavgsgmgtgqragspaagpplqpreplsysrlqrkslavdfvv 61 pslfrvyardlllppssselkagrpeargslaldcapllrllgpapgvswtagspapaea 121 rtlsrvlkggsvrklrrakqlvlelgeeailegovgppgeaavgllqfnlselfswwirq 181 gegrlrirlmpekkasevgregrlsaairasqprllfqifgtghsslesptnmpspspdy 241 ftwnitwimkdsfpflshrsryglecsfdfpceleyspplhdlrnqswswrripseeasq 301 mdlldgpgaerskemprgsf111ntsadskhtilspwmrsssehotlavsvhrhlqpsgr 361 yiaqllphneaareillmptpgkhgwtvlqgrigrpdnpfrvaleyissgnrslsavdff 421 alknosegtspgskmalqssftcwngtvlqlgqacdfhqdcaqgedesqmorklpvgfyc 481 nfedgfcgwtqgt1sphtpqwqvrtlkdarfqdhqdhalllsttdvpasesatvtsatfp 541 apiksspcelrmswlirgvlrgnvslvlvenktgkeqgrmvwhvaayeglslwqwmv1p1 601 ldvsdrfwlqmvawwgqgsraivafdnisisldcyltisgedkilqntapksrnlfernp 661 nkelkpgensprqtpifdptvhwlfttcgasgphgptqaqcnnayqnsnlsvevgsegpl 721 kgiqiwkvpatdtysisgygaaggkggkntmmrshgvsvlgifnlekddmlyilvgqqge 781 dacpstngliqkvcigennvieeeirvnrsvhewaggggggggatyvfkmkdgvpvplii 841 aaggggraygaktdtfhperlennssvlglngnsgaagggggwndntsllwagkslqega 901 tgghscpqamkkwgwetrggfggggggcssggggggyiggnaasnndpemdgedgvsfis 961 plgilytpalkvmeghgevnikhylncshcevdechmdpeshkvicfcdhgtvlaedgvs 1021 civsptpephlplslilsvvtsalvaalvlafsgimivyrrkhgelqamqmelqspeykl 1081 sklrtstimtdynpnycfagktssisdlkevprknitlirglghgafgevyegqvsgmpn 1141 dpsplqvavktlpevcseqdeldflmealiiskfnhqnivrcigvslqs1prfillelma 1201 ggdlksflretrprpsqpsslamldllhvardiacgcqyleenhfihrdiaarnolltcp 1261 gpgrvakigdfgmardiyrasyyrkggcamlpvkwmppeafmegiftsktdtwsfgvllw 1321 eifslgympypsksnqevlefvtsggrmdppknopgpvyrimtqcwqhqpedrpnfaiil 1381 erieyctqdpdvintalpieygplveeeekvpvrpkdpegvppllvsqqakreeerspaa 1441 ppplpttssgkaakkptaaeisvrvprgpavegghvnmafsqsnppselhkvhgsrnkpt 1501 slwnptygswftekptkknnpiakkephdrgnlglegsctvppnvatgrlpgasllleps 1561 sltanmkevplfrlrhfpcgnvnygyqqqglpleaatapgaghyedtilksknsmnqpgp ALK cDNA Sequence Reference SEQ ID NO: 7715 atgggagocatcgggctcctgtggct -,ctgccgctgctgctttccacggoagotgtgggc tccgggatggggaccggccagcgcgcgggctccccagctgr.ggggccgccgctgcagr.cc cgggagccactcagctactcgcgcctgcagaggaagagtctggcagttgacttcgtggtg ccctcgctcttccgtgtctacgcr.cgggacctactgotgccaccatcctcctcggagctg aaggotggcaggcccgaggcccgcggctcgctagctctggactgcgccccgctgr.tcagg ttgotggggccggcgccgggggt -,tcr.tggaccgccggt 7accagccccggoagaggcc cggacgctgt.ccagggtgotgaagggcggctccgtgcgcaagctccggcgtgocaagcag ttggtgctggagctgggcgaggaggcgatct tggagggt tgcgtcgggccccr.cggggag gcggotgtggggctgctccagtt atctcagogagctgttcagttggtggatt 7gccaa ggcgaagggcgactgaggatccgcctgatgoccgagaagaaggcgtcggaagtgggcaga gagggaaggctgtccgcggcaattcgcgcctcccagcr.ccgccttctcttccagatcttc EVZ
bbooeqeqbwobbbwoobqoeebee000e000ebbqebboobbebbqbeooeoqbwl bebbqoqqbeebbeooeeobeeeeobe0000eqeoobqeqeqebbqqoqoqwloqeeeb berleqobwbqbebbqqqooqbbqeoeoebeoeeeeqoqqoeoTIE-leebbeebbqeoT1 gg oobbebe00000bqebbqbeeqqbeoobwbqeoobqbqobbebbbeeebeqewewbe bobbbeoewqeoebeb000bbqebbboqwebebbTlebeeoobbqbebeebbwoobb eooqbwoebTloqoobqoeeebeoofrlobTleoebebooeooqeoqweooeeeebbeb bwleqbeo=lerloberlerloobTleoebbbowbbqboeobwqqooebbqobqeoobbqo oowowoobeoobeb000bowoob000ebebebooqooTlooqbeeowoebe55555 Og bobbqeowbebbqobwoqeoqqbb0000bwooqeeobwobeb.15555Tleobwbo TlerlqeoeebeooeooeeoTleeeobeoqeoqebwoobeebbqeowoqq=lebbweeb oebbeoeebqoqobqbqbeebwobwboebeebqbqobbqbeeobw0000bee000eb oee000bqeebbooqbqbbeoobbeefrIE-15.155e5555.1.1.1335355.1e33555.13.155 bboTleowooeoqeoeeeeebboboobqbbebbeebwoebqbeoqeoowowoebee gi7 oberlobqq=lobqoewee0000eeoewebooebqeoqeooebowoeoboowbeeobe bwbeeoeqbebwoobebeobwbebbqebeobqeoobeeobwbebbeooeobeebbo obooE-15.15Tlebqeoqeobbooqoqqqobbqooqbbwooboobbqbowoobqowoe erlberlerlow=looqefrloboqoqoeoobwoeoeoobebb0000e000eoqbqbTleobq ooqoqbobbqebbebqobbqobqbboebbboeooebqbqoqqobqoqeoqbbeeoeoobe 017 eebwooebbqeoeoqerleeboebeqbbebqbqoeoqbeobweeeqoqeTleobeeTle qeebqbeebbbboeoobbeebbqebqbeeeeqq=lobe0000eoeqbqooqeobbbweoo qbeoqeoTlooqqqbbberlebeebbbfrlebbqeeeb0000ebqeeoeeeowobeobqee obbobbeqeqeqebbebbobbebbebbqbbeowower1555555e55.155555ebboT1 .155555ebeeoebebbbqbbbberlbeebeebqeoobbe00000bwoTleoebbebbooe CE
oobqbbbebbeobqqqoqeeeebboobbbqoqobTlooqweoeeqebqeebbqobbqbb ebbqbbqbbeoboobebbooTleeobboeeewbbbewqqbboqooqoeeqeebebbqo ebebebe000eooTlboeoebeoebeeoobbbboewobbbeobbqbbqbbebboobeob Tleoqebw000bqbboobqbebbqebbeebqebee=ITIE-lboewoeoobebb.155555 ebbebbebbobbebbeobbbqbebqeobqbobeebeoeebqbqbooqeeebeebeebeqe OE
erlerleeoeebebebbTleobwqbeeebeooqeeqqbeooeeeoeqbe0000bwoboeb bebebbbeobeobbbqqbb=looqeoeqerlobqeoeerlebbeebebbwoeeoTloqeobb bwerlerloqbqbobboe000qbbobqebqeooeoeebeebbbobbeeebbbobbqobwb ebboewbbbowqeobeoewoeoebooeoobeoobqbbeebbqoqebeooqe355eee bw0000bbbebobebbbbqbbebbqbobebwoeeooweebeooewoboeeoeeobq cZ
beoeobbe000e0000bbqe0000bbbobeoobbberlerleoeooeoqqbqobbTleoqqb eoewooebqqqoqe00000ebeoebeeooeoTleeeebbbb000eeebwbebbeeoee e000eeebebebqqqbwoeeebeeoqeee000eobeoeqeebeobwoqebeeoebbeb ebbobeTleooeowoeqobwebbwobeoqeooqoqeqeeoebqwlobbqboqeoob ebeooqebbeeoebbberlberleoboqberlebeobwbbqoqqbbeoebqoqbqerlebow OZ
ow=loobTlerlberlebbqbeobbqbqoobebTlobbeebqewobooboqbqeobbqoqb bqebbeobbeeobebbeebbbooeeeeoeebebbqbeqobqbbqwoqbqboeeebbbbe bqqoqbeberlboTleowbbwoqb=leeboowbebqbqeooqowbebeeoqebooeob qooqq=lboewbqbeooebqbeoewbqbeeebqoqwb0000qbqebweooeqbeow 5.1.1E-loqobqeooebeeooeooebbeooqqbb000bqebbeeewooebbeoqbbeobbq gi eeowoweoe0000eoqbqoeoeobbee000ebbqobbqbqoqqobbqebeebqqqoee obwewITIbberlerloobweeebboobqbqebeoobebebqebeebebbbe000bqbq oebbeooeooqwebqbwobbeobbbTlobeoowoqbeoebbbqeebbqqbqqoeoT1 oowbebeobwoobbqebeeoowbbe0000qeoeebbeebqbeobweebeebwoob qq=loqwebbqbeobwqbqqobeobooeeebbqbeooqoqeoeqeebbwoobbqbebo OI
qq=le000eeoebeooqbobbboqeebeebbbeoowbqbeoebbqqbbqeobeebbbeoo we000bqebwowoqebebebeeobwbbeboeeoe0000bqobwbe000bTleoeq bbeeberlowoobeobwoeobbeoeobqbboqoqboobbqoeoeobweobebqbeobe obebbebqebbqbooqbebwoqeooeoeobeeoowebqobeowoeoeeowqqooqo qq=looqobbebe000frlebebbeeqoqqbobebeobbbbqoobbbqebbqobqwebbqe g be000wobbebbeboow000qeoboobobbwoqbbqobebeooeebbeowoebqeo bweoow000Tleqbebbqobebqbw000qwebqqqobeobqbebbqoq55.1E-lebo obeoboqeowqbwoqq=l000Tlooqoebeeeerleeqebbwoeow=leebbqeoewl qe=xleb=loo=loTloo=loTloob=leoeeeoeeooeo=leebbTlooqobeqeoqbbqoebbb 0S6I9ONTOZSI1/I3d f 19Zi=0/t 10Z OM
9Z-1:1-STOZ L6988ZO VD

WO 2(114/(152613 ataatgactcagtgctggcaacatcagcctgaagacaggcccaactttgccatcattttg gagaggattgaatactgcacccaggacccggatgtaatcaacaccgctttgccgatagaa tatggtccacttgtggaagaggaagagaaagtgcctgtgaggcccaaggaccctgagggg gttcctcctctcctggtctctcaacaggcaaaacgggaggaggagcgcagcccagctgcc ccaccacctctgcctaccacctcctctggcaaggctgcaaagaaacccacagctgcagag atctctgttcgagtccctagagggccggccgtggaagggggacacgtgaatatggcattc tctcagtccaaccctccttcggagttgcacaaggtccacggatccagaaacaagcccacc agcttgtggaacccaacgtacggctcctggtttacagagaaacccaccaaaaagaataat cctatagcaaagaaggagccacacgacaggggtaacctggggctggagggaagctgtact gtcccacctaacgttgcaactgggagacttccgggggcctcactgctcctagagccctct tcgctgactgccaatatgaaggaggtacctctgttcaggctacgtcacttcccttgtggg aatgtcaattacggctaccagcaacagggcttgcccttagaagccgctactgcccctgga gctggtcattacgaggataccattctgaaaagcaagaatagcatgaaccagcctgggccc tga EGFR cDNA Sequence Reference SEQ ID NO: 7716 ATGCGACCCTCCGGGACGGCCGGGGCAGCGCTCCTGGCGCTGCTGGCTGCGCTCTGCCCG
GCGAGTCGGGCTCTGGAGGAAAAGAAAGTTTGCCAAGGCACGAGTAACAAGCTCACGCAG
TTGGGCACTTTTGAAGATCATTTTCTCAGCCTCCAGAGGATGTTCAATAACTGTGAGGTG
GTCCTTGGGAATTTGGAAATTACCTATGTGCAGAGGAATTATGATCTTTCCTTCTTAAAG
ACCATCCAGGAGGTGGCTGGTTATGTCCTCATTGCCCTCAACACAGTGGAGCGAATTCCT
TTGGAAAACCTGCAGATCATCAGAGGAAATATGTACTACGAAAATTCCTATGCCTTAGCA
GTCTTATCTAACTATGATGCAAATAAAACCGGACTGAAGGAGCTGCCCATGAGAAATTTA
CAGGAAATCCTGCATGGCGCCGTGCGGTTCAGCAACAACCCTGCCCTGTGCAACGTGGAG
AGCATCCAGTGGCGGGACATAGTCAGCAGTGACTTTCTCAGCAACATGTCGATGGACTTC
CAGAACCACCTGGGCAGCTGCCAAAAGTGTGATCCAAGCTGTCCCAATGGGAGCTGCTGG
GGTGCAGGAGAGGAGAACTGCCAGAAACTGACCAAAATCATCTGTGCCCAGCAGTGCTCC
GGGCGCTGCCGTGGCAAGTCCCCCAGTGACTGCTGCCACAACCAGTGTGCTGCAGGCTGC
ACAGGCCCCCGGGAGAGCGACTGCCTGGTCTGCCGCAAATTCCGAGACGAAGCCACGTGC
AAGGACACCTGCCCCCCACTCATGCTCTACAACCCCACCACGTACCAGATGGATGTGAAC
CCCGAGGGCAAATACAGCTTTGGTGCCACCTGCGTGAAGAAGTGTCCCCGTAATTATGTG
GTGACAGATCACGGCTCGTGCGTCCGAGCCTGTGGGGCCGACAGCTATGAGATGGAGGAA
GACGGCGTCCGCAAGTGTAAGAAGTGCGAAGGGCCTTGCCGCAAAGTGTGTAACGGAATA
GGTATTGGTGAATTTAAAGACTCACTCTCCATAAATGCTACGAATATTAAACACTTCAAA
AACTGCACCTCCATCAGTGGCGATCTCCACATCCTGCCGGTGGCATTTAGGGGTGACTCC
TTCACACATACTCCTCCTCTGGATCCACAGGAACTGGATATTCTGAAAACCGTAAAGGAA
ATCACAGGGTTTTTGCTGATTCAGGCTTGGCCTGAAAACAGGACGGACCTCCATGCCTTT
GAGAACCTAGAAATCATACGCGGCAGGACCAAGCAACATGGTCAGTTTTCTCTTGCAGTC
GTCAGCCTGAACATAACATCCTTGGGATTACGCTCCCTCAAGGAGATAAGTGATGGAGAT
GTGATAATTTCAGGAAACAAAAATTTGTGCTATGCAAATACAATAAACTGGAAAAAACTG
TTTGGGACCTCCGGTCAGAAAACCAAAATTATAAGCAACAGAGGTGAAAACAGCTGCAAG
GCCACAGGCCAGGTCTGCCATGCCTTGTGCTCCCCCGAGGGCTGCTGGGGCCCGGAGCCC
AGGGACTGCGTCTCTTGCCGGAATGTCAGCCGAGGCAGGGAATGCGTGGACAAGTGCAAC
CTTCTGGAGGGTGAGCCAAGGGAGTTTGTGGAGAACTCTGAGTGCATACAGTGCCACCCA
GAGTGCCTGCCTCAGGCCATGAACATCACCTGCACAGGACGGGGACCAGACAACTGTATC
CAGTGTGCCCACTACATTGACGGCCCCCACTGCGTCAAGACCTGCCCGGCAGGAGTCATG
GGAGAAAACAACACCCTGGTCTGGAAGTACGCAGACGCCGGCCATGTGTGCCACCTGTGC
CATCCAAACTGCACCTACGGATGCACTGGGCCAGGTCTTGAAGGCTGTCCAACGAATGGG
CCTAAGATCCCGTCCATCGCCACTGGGATGGTGGGGGCCCTCCTCTTGCTGCTGGTGGTG
GCCCTGGGGATCGGCCTCTTCATGCGAAGGCGCCACATCGTTCGGAAGCGCACGCTGCGG
AGGCTGCTGCAGGAGAGGGAGCTTGTGGAGCCTCTTACACCCAGTGGAGAAGCTCCCAAC
CAAGCTCTCTTGAGGATCTTGAAGGAAACTGAATTCAAAAAGATCAAAGTGCTGGGCTCC
GGTGCGTTCGGCACGGTGTATAAGGGACTCTGGATCCCAGAAGGTGAGAAAGTTAAAATT
CCCGTCGCTATCAAGGAATTAAGAGAAGCAACATCTCCGAAAGCCAACAAGGAAATCCTC
GATGAAGCCTACGTGATGGCCAGCGTGGACAACCCCCACGTGTGCCGCCTGCTGGGCATC

WO 2(114/(152613 TGCCTCACCTCCACCGTGCAGCTCATCACGCAGCTCATGCCCTTCGGCTGCCTCCTGGAC
TATGTCCGGGAACACAAAGACAATATTGGCTCCCAGTACCTGCTCAACTGGTGTGTGCAG
ATCGCAAAGGGCATGAACTACTTGGAGGACCGTCGCTTGGTGCACCGCGACCTGGCAGCC
AGGAACGTACTGGTGAAAACACCGCAGCATGTCAAGATCACAGATTTTGGGCTGGCCAAA
CTGCTGGGTGCGGAAGAGAAAGAATACCATGCAGAAGGAGGCAAAGTGCCTATCAAGTGG
ATGGCATTGGAATCAATTTTACACAGAATCTATACCCACCAGAGTGATGTCTGGAGCTAC
GGGGTGACTGTTTGGGAGTTGATGACCTTTGGATCCAAGCCATATGACGGAATCCCTGCC
AGCGAGATCTCCTCCATCCTGGAGAAAGGAGAACGCCTCCCTCAGCCACCCATATGTACC
ATCGATGTCTACATGATCATGGTCAAGTGCTGGATGATAGACGCAGATAGTCGCCCAAAG
TTCCGTGAGTTGATCATCGAATTCTCCAAAATGGCCCGAGACCCCCAGCGCTACCTTGTC
ATTCAGGGGGATGAAAGAATGCATTTGCCAAGTCCTACAGACTCCAACTTCTACCGTGCC
CTGATGGATGAAGAAGACATGGACGACGTGGTGGATGCCGACGAGTACCTCATCCCACAG
CAGGGCTTCTTCAGCAGCCCCTCCACGTCACGGACTCCCCTCCTGAGCTCTCTGAGTGCA
ACCAGCAACAATTCCACCGTGGCTTGCATTGATAGAAATGGGCTGCAAAGCTGTCCCATC
AAGGAAGACAGCTTCTTGCAGCGATACAGCTCAGACCCCACAGGCGCCTTGACTGAGGAC
AGCATAGACGACACCTTCCTCCCAGTGCCTGAATACATAAACCAGTCCGTTCCCAAAAGG
CCCGCTGGCTCTGTGCAGAATCCTGTCTATCACAATCAGCCTCTGAACCCCGCGCCCAGC
AGAGACCCACACTACCAGGACCCCCACAGCACTGCAGTGGGCAACCCCGAGTATCTCAAC
ACTGTCCAGCCCACCTGTGTCAACAGCACATTCGACAGCCCTGCCCACTGGGCCCAGAAA
GGCAGCCACCAAATTAGCCTGGACAACCCTGACTACCAGCAGGACTTCTTTCCCAAGGAA
GCCAAGCCAAATGGCATCTTTAAGGGCTCCACAGCTGAAAATGCAGAATACCTAAGGGTC
GCGCCACAAAGCAGTGAATTTATTGGAGCATGA
BRAF cDNA Sequence Reference SEQ ID NO: 7717 atggcggcgctgagcggtggcggtggtggcggcgcggagccgggccaggctctgttcaac ggggacatggagcccgaggccggcgccggcgccggcgccgcggcctcttcggctgcggac cctgccattccggaggaggtgtggaatatcaaacaaatgattaagttgacacaggaacat atagaggccctattggacaaatttggtggggagcataatccaccatcaatatatctggag gcctatgaagaatacaccagcaagctagatgcactccaacaaagagaacaacagttattg gaatctctggggaacggaactgatttttctgtttctagctctgcatcaatggataccgtt acatcttcttcctcttctagcctttcagtgctaccttcatctctttcagtttttcaaaat cccacagatgtggcacggagcaaccccaagtcaccacaaaaacctatcgttagagtcttc ctgcccaacaaacagaggacagtggtacctgcaaggtgtggagttacagtccgagacagt ctaaagaaagcactgatgatgagaggtctaatcccagagtgctgtgctgtttacagaatt caggatggagagaagaaaccaattggttgggacactgatatttcctggcttactggagaa gaattgcatgtggaagtgttggagaatgttccacttacaacacacaactttgtacgaaaa acgtttttcaccttagcattttgtgacttttgtcgaaagctgcttttccagggtttccgc tgtcaaacatgtggttataaatttcaccagcgttgtagtacagaagttccactgatgtgt gttaattatgaccaacttgatttgctgtttgtctccaagttctttgaacaccacccaata ccacaggaagaggcgtccttagcagagactgccctaacatctggatcatccccttccgca cccgcctcggactctattgggccccaaattctcaccagtccgtctccttcaaaatccatt ccaattccacagcccttccgaccagcagatgaagatcatcgaaatcaatttgggcaacga gaccgatcctcatcagctcccaatgtgcatataaacacaatagaacctgtcaatattgat gacttgattagagaccaaggatttcgtggtgatggaggatcaaccacaggtttgtctgct accccccctgcctcattacctggctcactaactaacgtgaaagccttacagaaatctcca ggacctcagcgagaaaggaagtcatcttcatcctcagaagacaggaatcgaatgaaaaca cttggtagacgggactcgagtgatgattgggagattcctgatgggcagattacagtggga caaagaattggatctggatcatttggaacagtctacaagggaaagtggcatggtgatgtg gcagtgaaaatgttgaatgtgacagcacctacacctcagcagttacaagccttcaaaaat gaagtaggagtactcaggaaaacacgacatgtgaatatcctactcttcatgggctattcc acaaagccacaactggctattgttacccagtggtgtgagggctccagcttgtatcaccat ctccatatcattgagaccaaatttgagatgatcaaacttatagatattgcacgacagact gcacagggcatggattacttacacgccaagtcaatcatccacagagacctcaagagtaat aatatatttcttcatgaagacctcacagtaaaaataggtgattttggtctagctacagtg aaatctcgatggagtgggtcccatcagtttgaacagttgtctggatccattttgtggatg WO 2(114/(152613 gcaccagaagtcatcagaatgcaagataaaaatccatacagctttcagtcagatgtatat gcatttggaattgttctgtatgaattgatgactggacagttaccttattcaaacatcaac aacagggaccagataatttttatggtgggacgaggatacctgtctccagatctcagtaag gtacggagtaactgtccaaaagccatgaagagattaatggcagagtgcctcaaaaagaaa agagatgagagaccactctttccccaaattctcgcctctattgagctgctggcccgctca ttgccaaaaattcaccgcagtgcatcagaaccctccttgaatcgggctggtttccaaaca gaggattttagtctatatgcttgtgcttctccaaaaacacccatccaggcagggggatat ggtgcgtttcctgtccactga KRAS cDNA Sequence Reference SEQ ID NO: 7718 atgactgaatataaacttgtggtagttggagctggtggcgtaggcaagagtgccttgacg atacagctaattcagaatcattttgtggacgaatatgatccaacaatagaggattcctac aggaagcaagtagtaattgatggagaaacctgtctcttggatattctcgacacagcaggt caagaggagtacagtgcaatgagggaccagtacatgaggactggggagggctttctttgt gtatttgccataaataatactaaatcatttgaagatattcaccattatagagaacaaatt aaaagagttaaggactctgaagatgtacctatggtcctagtaggaaataaatgtgatttg ccttctagaacagtagacacaaaacaggctcaggacttagcaagaagttatggaattcct tttattgaaacatcagcaaagacaagacagggtgttgatgatgccttctatacattagtt cgagaaattcgaaaacataaagaaaagatgagcaaagatggtaaaaagaagaaaaagaag tcaaagacaaagtgtgtaattatgtaa

Claims (47)

What is claimed is:

1. A kinase inhibitor resistance panel comprising one or more primer sets from one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT .

2. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS.

3. The kinase inhibitor panel of claim 2, wherein one or more KRAS
hybridizing primers or primer sets comprise one or more of the primers of Tables 10 and/or 14.

4. The kinase inhibitor panel of claim 2, wherein one or more KRAS
hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 4601-5200 and 7181-7610.

5. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 18, 19, 20, 21 or 22 of EGFR.

6. The kinase inhibitor panel of claim 5, wherein one or more EGFR
hybridizing primers or primer sets comprise one or more of the primers of Tables 8 and/or 12.

7. The kinase inhibitor panel of claim 6, wherein one or more EGFR
hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1641-2440 and 5819-6524.

8. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 21, 22, 23, 24, or 25 of ALK.

9. The kinase inhibitor panel of claim 8, wherein one or more ALK
hybridizing primers or primer sets comprise one or more of the primers of Tables 7 and/or 11.

10. The kinase inhibitor panel of claim 9, wherein one or more ALK
hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1-1640 and 5201-5818.

11. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 8, 9, 10, 11, 12, 13, or 17 of KIT.

12. The kinase inhibitor panel of claim 11, wherein one or more KIT
hybridizing primers or primer sets comprise one or more of the primers of Table 9.

13. The kinase inhibitor panel of claim 12, wherein one or more KIT
hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 2441-4600.

14. The kinase inhibitor resistance panel of claim 1, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 10, 11, 13, 14, or 15 of BRAF.

15. The kinase inhibitor panel of claim 14, wherein one or more BRAF
hybridizing primers or primer sets comprise one or more of the primers of Table 13.

16. The kinase inhibitor panel of claim 15, wherein one or more BRAF
hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 6525-7180.

17. The kinase inhibitor panel of claim 1, wherein the panel comprises two or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

18. The kinase inhibitor panel of claim 1, wherein the panel comprises 3, 4, 5, 6, 7, 8, 9, 10, or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

19. The kinase inhibitor panel of claim 1, wherein the panel comprises one or more primer sets for 2, 3, 4, of all 5 of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

20. The kinase inhibitor of claim 1, wherein the kinase inhibitor resistance is resistance to an ALK kinase inhibitor.

21. The kinase inhibitor resistance panel of claim 20, wherein the kinase inhibitor is selected from the group consisting of crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib.

22. The kinase inhibitor resistance panel of claim 21, wherein the kinase inhibitor comprises crizotinib.

23. The kinase inhibitor panel of claim 1, wherein the one or more primer sets amplify one or more mutations associated with kinase inhibitor resistance, wherein the mutation is identified in Table 2, 3, 4, 5, or 6.

24. A method for the detection of kinase inhibitor resistance comprising obtaining a tissue sample from a subject with a cancer and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample using the kinase inhibitor resistant panel of Claim 1, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.

25. A method for the detection of kinase inhibitor resistance comprising obtaining a tissue sample from a subject with a cancer and conducting a high throughput sequencing (also known as next generation sequencing) reaction on the sample using one or more primer sets or primer panels with primer sets that specifically hybridizes to one or more of the genes selected from the group consisting of ALK, KRAS, EGFR, KIT, and BRAF, wherein the presence of a mutation in the nucleic acid sequence of a gene associated with kinase inhibitor resistance indicates that that the cancer is resistant or will become resistant to a kinase inhibitor.

26. The method of Claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 1 or 2 of KRAS.

27. The method of claim 26, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of Tables 10 and/or 14.

28. The method of claim 26, wherein one or more KRAS hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 4601-5200 and 7181-7610.

29. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 18, 19, 20, 21 or 22 of EGFR.

30. The method of claim 29, wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of Tables 8 and/or 12.

31. The method of claim 29, wherein one or more EGFR hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1641-2440 and 5819-6524.

32. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 21, 22, 23, 24, or 25 of ALK.

33. The method of claim 32, wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of Tables 7 and/or 11.

34. The method of claim 32, wherein one or more ALK hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 1-1640 and 5201-5818.

35. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 8, 9, 10, 11, 12, 13, or 17 of KIT.

36. The method of claim 35, wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of Table 9.

37. The method of claim 35, wherein one or more KIT hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 2441-4600.

38. The method of claim 25, wherein at least one primer sets hybridizes and amplifies nucleic acid from exon 10, 11, 13, 14, or 15 of BRAF .

39. The method of claim 38, wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of Table 13.

40. The method of claim 38, wherein one or more BRAF hybridizing primers or primer sets comprise one or more of the primers of SEQ ID NOs: 6525-7180.

41. The method of claim 25, wherein the panel comprises two or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

42. The method of claim 25, wherein the panel comprises 3, 4, 5, 6, 7, 8, 9, 10, or more primer sets for one or more of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

43. The method of claim 25, wherein the panel comprises one or more primer sets for 2, 3, 4, of all 5 of the genes selected from group of genes comprising KRAS, BRAF, EGFR, ALK, and KIT.

44. The method of claim 25, wherein the kinase inhibitor resistance is resistance to an ALK kinase inhibitor.

45. The method of claim 25, wherein the kinase inhibitor is selected from the group consisting of crizotinib, afatinib, Axitinib, bevacizumab, Bosutinib, Cetuximab, Dasatinib, Erlotinib, Fostamati nib, Gefitinib, Imatinib, Lapatinib, Lenvatinib, Nilotinib, Panitumumab, Pazopanib, Pegaptanib, Ranibizumab, Ruxolitinib, Sorafenib, Sunitinib, Trastuzumab, and Vemurafenib.

46. The method of claim 45, wherein the kinase inhibitor comprises crizotinib.

47. The method of claim 25, wherein the one or more primer sets amplify one or more mutations associated with kinase inhibitor resistance, wherein the mutation is identified in Table 2, 3, 4, 5, or 6.