CN111575378A - Detection method, detection composition and detection kit for hereditary breast cancer and ovarian cancer syndrome - Google Patents
Detection method, detection composition and detection kit for hereditary breast cancer and ovarian cancer syndrome Download PDFInfo
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Abstract
The invention relates to a detection method, a detection composition and a detection kit for hereditary breast cancer and ovarian cancer syndromes, wherein a BRCA1 gene coding region and a plurality of SNP sites on the upstream and downstream of a BRCA1 gene are obtained by screening aiming at the hereditary breast cancer and ovarian cancer syndromes, and a genetic detection method before embryo implantation with strong universality, high diagnosis rate and low cost can be established on a second-generation sequencing platform based on the coding region and the plurality of SNP sites on the upstream and downstream. The embryo detection result can be determined by detecting the sequence information of the coding region and the upstream and downstream SNP sites of the detected sample and carrying out analysis and comparison. Besides the coding region, 157 SNPs are selected for analysis on the upstream and downstream of the BRCA1 gene, the universality is very high, a large number of samples can be analyzed at one time, the result reliability is high, and the detection cost is low.
Description
Technical Field
The invention relates to the field of gene detection, in particular to a detection method, a detection composition and a detection kit for hereditary breast cancer and ovarian cancer syndromes.
Background
Breast cancer (BRC) is one of the most common malignancies in women. Most breast cancers are sporadic, and some breast cancers are genetically predisposed, manifested as familial aggregation, and are mostly less than 50 years old. Breast cancer susceptibility gene 1 (BRCA 1) and BRCA2 gene mutations cause about 60% of Hereditary Breast cancers and are the only factors currently known for Hereditary Breast cancer and ovarian cancer syndrome (HBOC). Compared with the general population, the carrier of BRCA1 and BRCA2 mutations has obviously increased risk of the breast cancer and ovarian cancer in the whole life, and has higher risk of the breast cancer or the ovarian cancer.
Hereditary breast and ovarian cancer syndromes are autosomal diseases with carriers at 50% risk of transmitting mutations to their offspring. The prenatal diagnosis or the genetic testing (PGT) before embryo implantation of female carriers is an effective means for blocking the transmission of breast cancer and ovarian cancer, eliminates high-risk pathogenic factors in families and reduces the incidence rate of tumors. PGT refers to the process of when the embryo develops in vitro to the cleavage stage or the blastocyst stage, several cells are biopsied for genetic detection, and finally the embryo without the risk of disease is selected to be implanted into the mother uterus, thereby achieving the purpose of producing healthy offspring. PGT can effectively avoid physical and psychological damage caused by terminating pregnancy due to pregnancy genetic disease fetus, and becomes the first choice for fetus couples with high risk of birth genetic disease. Therefore, how to predict BRCA mutation condition of embryo more accurately is crucial, so that corresponding preventive measures are taken.
Disclosure of Invention
Therefore, the detection method, the detection composition and the kit for the hereditary breast cancer and ovarian cancer syndrome before embryo implantation have the advantages of strong universality, high diagnosis rate and low cost.
A method for detecting hereditary breast cancer and ovarian cancer syndrome before embryo implantation, comprising the following steps:
obtaining genome DNA of an embryo;
detecting sequence information of a BRCA gene coding region and SNP sites on the upper and lower reaches of the BRCA gene in the genome DNA, wherein the SNP sites on the BRCA gene coding region and the upper and lower reaches of the BRCA gene comprise chr, chr, The composition is used for preventing and treating the diseases of the liver, the kidney, the liver, the spleen, the kidney, the liver, the kidney, the spleen, the kidney, the liver, the kidney, the liver, the kidney, the liver, the, The composition is used for treating the diseases of the liver, the kidney, the spleen, the stomach, the spleen, the kidney, the spleen and the kidney, wherein the chr is, the chr is, the chr, the chor, the CHr;
and (c) analyzing the sequence information in alignment to determine whether the embryo has hereditary breast cancer and ovarian cancer syndrome.
In one embodiment, the step of detecting the sequence information comprises the steps of: and carrying out multiplex PCR on the genome DNA by using a plurality of pairs of primers capable of respectively and specifically amplifying the coding region of the BRCA1 gene and the SNP sites at the upstream and downstream of the BRCA1 gene to obtain an amplification product, and sequencing the amplification product to obtain the sequence information.
In one embodiment, the primer is selected from SEQ ID NO: 1 to SEQ ID NO: 338.
In one embodiment, the step of performing alignment analysis comprises the steps of: and comparing the sequence information with DNA sequences of both parents, and analyzing the haplotype of the embryo.
In one embodiment, the step of performing alignment analysis further comprises the steps of: and comparing the sequence information with a human genome reference sequence, and analyzing the SNP coverage multiple and the genotype.
In one embodiment, the step of obtaining the genomic DNA comprises the steps of: trophoblast cells are collected from embryos that develop to the cleavage or blastocyst stage and the DNA in the trophoblast cells is subjected to whole genome amplification.
A detection composition for detecting hereditary breast cancer and ovarian cancer syndrome, comprising a plurality of detection agents capable of detecting sequence information of the coding region of BRCA1 gene and SNP sites upstream and downstream of BRCA1 gene, respectively.
In one embodiment, the detection agent is a PCR primer, and the detection agent is selected from SEQ ID NO: 1 to SEQ ID NO: 338.
A detection kit for detecting hereditary breast cancer and ovarian cancer syndromes comprises the detection composition and a reaction reagent for library construction.
In one embodiment, the reaction reagents are selected from one or more of multiplex PCR polymerase, DNA ligase, end-repair enzymes, dNTPs, and PCR buffer.
The invention screens and obtains a BRCA1 gene coding region and a plurality of SNP sites on the upstream and downstream of a BRCA1 gene aiming at hereditary breast cancer and ovarian cancer syndromes, and a genetic detection method before embryo implantation with strong universality, high diagnosis rate and low cost can be established on a next-generation sequencing platform based on the coding region and the plurality of SNP sites on the upstream and downstream. The detection result of the hereditary breast cancer and ovarian cancer syndrome of the embryo can be determined by detecting the sequence information of the coding region and the upstream and downstream SNP sites of the detected sample and carrying out analysis and comparison. In addition to a coding region, 157 SNPs are selected from the upstream and downstream of the BRCA1 gene for analysis, the universality is very high, a cell level pre-experiment is omitted, a large number of samples can be analyzed at one time based on a high-throughput sequencing technology, the average sequencing depth can reach more than 100X, the result reliability is high, and the detection cost is low.
Detailed Description
In order that the invention may be more fully understood, a more particular description of the invention will now be rendered by reference to specific embodiments thereof that are illustrated in the appended drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
In the present invention, reads (reads) refer to sequence fragments obtained by sequencing; single Nucleotide Polymorphism (SNP) mainly refers to DNA sequence polymorphism caused by variation of a single nucleotide at the genome level; haplotypes (haplotypes) refer to a set of interrelated sets of single nucleotide polymorphisms, also known as haplotypes or haplotypes, located in specific regions of a chromosome that are predisposed to progeny in an overall inheritance; the sequencing depth, i.e., for example, in one embodiment, the sequencing depth is 1000X, which means that the strip of specific PCR amplification product is sequenced 1000 times.
The method for detecting hereditary breast cancer and ovarian cancer syndrome before embryo implantation in one embodiment of the invention comprises the following steps of S1-S3:
s1, obtaining genome DNA of the embryo.
S, detecting sequence information of a BRCA gene coding region and SNP sites on the upper and lower reaches of the BRCA gene in genomic DNA, wherein the SNP sites on the BRCA gene coding region and the upper and lower reaches of the BRCA gene comprise chr, chr, The composition is used for preventing and treating the diseases of the liver, the kidney, the liver, the spleen, the kidney, the liver, the kidney, the spleen, the kidney, the liver, the kidney, the liver, the kidney, the liver, the, chr, chr.
S3, and comparing and analyzing the sequence information to determine whether the embryo has hereditary breast cancer and ovarian cancer syndrome.
According to the european human reproductive and embryonic association (ESHRE) "guidelines for amplification-based PGD practice" (2011 edition), pre-implantation genetic testing of monogenic genetic diseases is proposed, using a strategy of mutation detection combined with haplotype analysis to ensure the accuracy of embryo test results. Common methods for performing gene detection at the embryo level are multiplex nested PCR and Karyomapping techniques. The multiple nested PCR method needs to screen heterozygous short tandem repeat Sequences (STR) for BRCA1 female carriers in advance, then establishes a multiple PCR system containing mutation sites and heterozygous STR sites at lymphocyte level, evaluates the amplification efficiency and the allele tripping rate of each site at cell level, and can be applied to embryo detection after qualification. Due to the limited number of STRs and different heterozygosity frequencies in the population, the design of the multiple nested PCR method is more personalized and has lower universality. The strong individuation also means large workload and long detection period.
Compared with STR sites, the number of single nucleotide polymorphic Sites (SNP) is large and wide in distribution, the occurrence frequency in the population exceeds 1%, the total number of SNP is 300 ten thousand, the average number of SNP is 1/3 Kb, and the automatic analysis is easy to realize on a high-throughput sequencing platform. The Karyomapping technology is used for indirectly eliminating gene defects through SNP linkage analysis. The chip is provided with 30 ten thousand SNP probes for linkage analysis of the whole genome, and is a comprehensive and universal PGD technology for the monogenic diseases. However, Karyomapping cannot detect gene mutations, so proband samples or appropriate family member samples are necessary. If there are not enough pedigree samples, additional mutation detection is required. Therefore, there is a need to develop a method for detecting all coding regions of BRCA1 gene and covering enough SNP sites in and upstream and downstream of the gene, which is used for determining embryo genotype.
The BRCA1 gene is located at 17q21, has a full length of 100kb, contains 22 coding exons, 2 non-coding exons and 22 introns, has a larger exon 11, codes 61% of amino acid residues, transcribes 7.8kb of mRNA from 22 exons, and has a protein containing 1863 amino acid residues, a molecular weight of 220KD and contains Alu repetitive sequences up to 41.5% and other repetitive sequences up to 4.8%. The N-terminal sequence of the protein encoded by BRCA1 contains a loop domain (ringdomain) capable of forming a loop 2 heterodimer with a BRCA1 related loop protein (BRCA1 associated RING domain protein, BARD 1). The invention screens and obtains a BRCA1 gene coding region and a plurality of SNP sites on the upstream and downstream of a BRCA1 gene aiming at hereditary breast cancer and ovarian cancer syndromes, and a genetic detection method before embryo implantation with strong universality, high diagnosis rate and low cost can be established on a next-generation sequencing platform based on the coding region and the plurality of SNP sites on the upstream and downstream. The detection result of the hereditary breast cancer and ovarian cancer syndrome of the embryo can be determined by detecting the sequence information of the coding region and the upstream and downstream SNP sites of the detected sample and carrying out analysis and comparison. In addition to a coding region, 157 SNPs are selected from the upstream and downstream of the BRCA1 gene for analysis, the universality is very high, a cell level pre-experiment is omitted, a large number of samples can be analyzed at one time based on a high-throughput sequencing technology, the average sequencing depth can reach more than 100X, the result reliability is high, and the detection cost is low.
It is understood that the detection object of the detection method is an embryo which is not implanted into the uterus, and the detection result does not relate to the disease diagnosis result of both parents, and thus the detection method does not belong to a disease diagnosis and treatment method.
The SNP locus is finally determined based on the following screening principle and combined with long-term test optimization:
(1) high frequency SNP selected from genome project database of thousand people
(http://www.ncbi.nlm.nih.gov/variation/tools/1000genomes/)；
(2) High-frequency SNP loci with minimum allele frequencies greater than 0.2;
(3) removing the polynucleotide (polyN) and SNP sites with GC content of more than 70 percent in the sequence of 50bp upstream and downstream of the sites;
(4) removing SNP sites with multiple positions aligned to hg19 of a human genome by 50bp sequences (namely removing SNP sites with high homology);
(5) SNP selection range: SNP sites within 1Mb of the upstream and downstream of the gene are preferably selected, and if the number of SNPs within 1Mb is small, the range can be expanded appropriately, for example, 2Mb for each of the upstream and downstream.
In a specific example, the step of detecting the sequence information includes the steps of: and performing multiplex PCR on the genome DNA by using a plurality of pairs of primers capable of respectively and specifically amplifying the coding region of the BRCA1 gene and the SNP sites at the upstream and downstream of the BRCA1 gene to obtain an amplification product, and sequencing the amplification product to obtain sequence information. It is to be understood that the method of detecting the above sequence information is not limited thereto, and may be selected as needed by those skilled in the art.
In one specific example, the primer is selected from SEQ ID NO: 1 to SEQ ID NO: 338. Preferably, the primer comprises SEQ ID NO: 1 to SEQ ID NO: 338. the primers obtained by screening in the invention have high specificity and similar annealing temperature, and the sizes of PCR product fragments are all in the range of 125 bp-275 bp. The positions of coding regions and SNP sites on the chr17 upstream and downstream and the sequences corresponding to 169 primer pairs are shown in Table 1, wherein the sequence numbers 1-89 are the SNP sites and primer pairs within 2M downstream of the BRCA1 gene, the sequence numbers 90-101 are the SNP sites and primer pairs within 2M upstream of the BRCA1 gene, and the sequence numbers 102-169 are the SNP sites and primer pairs within 2M upstream of the BRCA1 gene. The coding region of BRCA1 gene and upstream and downstream SNP sites of the sample to be tested are amplified by mixing 169 pairs of primers into a PCR reaction tube and carrying out 169 reactions.
TABLE 1 SNP sites and primer sequence Listing
In a specific example, the step of obtaining genomic DNA comprises the steps of: trophoblast cells are collected from embryos that develop to the cleavage or blastocyst stage and the DNA in the trophoblast cells is subjected to whole genome amplification.
In a specific example, the step of performing the alignment analysis includes the following steps: the sequence information is compared with the DNA sequences of both parents to analyze the haplotype of the embryo.
In a specific example, the step of performing the alignment analysis further includes the steps of: and comparing the sequence information with a human genome reference sequence, and analyzing the SNP coverage multiple and the genotype. It is understood that the human genome reference sequence may be from a public database. Specifically, the human genome reference sequence may be a human genome reference sequence in NCBI or UCSC databases, such as hg19, hg38, and the like.
It is understood that the DNA sequences of both parents can also be obtained by sequencing DNA samples of both parents. Optionally, the DNA samples of both parents are selected from one or more of peripheral blood genome DNA, semen DNA, oral mucosa cell DNA and cell whole genome amplification products. Preferably, the amount of DNA in each DNA sample is greater than 500 ng.
In a specific example, the sequencing method is Ion Torrent PGM or Illumina Miseq, and library building is performed according to a corresponding standard library building flow. Sequence alignment during sequencing analysis can be achieved by aligning the reads to the reference genomic sequence using any sequence alignment program, such as BWA (Burrow-Wheeler-Aligner) available to those skilled in the art, to obtain the position of the reads on the reference genome. During data analysis, original data generated by a sequencer such as Illumina and the like can be subjected to linker sequence removal by utilizing trimmatic software, BWA software is compared to a human reference genome, and finally, haplotype SNP site coverage multiple and genotype are analyzed.
The detection composition for detecting hereditary breast cancer and ovarian cancer syndrome according to an embodiment of the present invention includes a plurality of detection agents capable of detecting sequence information of the coding region of BRCA1 gene and SNP sites upstream and downstream of BRCA1 gene, respectively.
Optionally, the detection agent is a PCR primer and is selected from SEQ ID NO: 1 to SEQ ID NO: 338. It is to be understood that the detecting agent is not limited thereto, and may be, for example, a probe or the like.
The detection kit for detecting hereditary breast cancer and ovarian cancer syndromes of one embodiment of the invention comprises the detection composition and a reaction reagent for library construction.
In one specific example, the reaction reagents for library construction are selected from one or more of multiplex PCR polymerase, DNA ligase, end-repair enzymes, dNTPs and PCR buffer. It is to be understood that the kit is not limited thereto, and various reagents may be added or subtracted as necessary.
In one specific example, the detection kit further comprises a tag sequence, and when the DNA molecule to be detected comes from a plurality of samples to be tested, each sample can be added with a different tag sequence (barcode) for sample differentiation during sequencing, thereby realizing simultaneous sequencing of a plurality of samples.
The following are specific examples.
Example 1
One woman and her mother had a c.4065_4068delTCAA heterozygous mutation in BRCA1 gene (NM _007294.3), and her mother was deceased by ovarian cancer. The female application performs a pre-embryo implantation genetic test (PGT) for the above-mentioned pathogenic mutations, yielding healthy offspring.
The female obtains 7 embryos (P917-JJJ-1-P917-JJJ-7) through PGT assisted pregnancy, the embryos develop to the blastocyst stage to carry out trophoblast cell biopsy, and the known variation and haplotype of BRCA1 gene are further detected after the biopsy cells are amplified by a whole genome.
Of 7 embryos, 5 were normal and 2 were carriers of maternal mutations. The couple transplants a normal embryo, carries out prenatal ultrasonic diagnosis and gene diagnosis in 16 weeks of pregnancy, B ultrasonic indicates that the fetus is normally developed, genetic detection indicates that the BRCA1 gene of the fetus does not have c.4065_4068delTCAA mutation, the result is consistent with the PGT result, the later pregnancy is successful, and a healthy child without BRCA1 gene c.4065_4068delTCAA is born.
The following is the pre-implantation genetic testing procedure:
1. library construction and sequencing
The PCR primers (SEQ ID NO: 1-SEQ ID NO: 338) are used for building a library of the biopsy cell whole genome amplification product according to the Illumina standard library building process, and a Miseq sequencer is used for sequencing.
2. Comparison and statistics
Removing a linker sequence from raw data generated by an Illumina sequencer by using trimmatic software, aligning the raw data to a human hg19 reference genome by using BWA software, and finally analyzing haplotype SNP coverage fold and genotype.
3. Analysis of results
The detection results are shown in table 2, wherein F0 and F1 represent male normal chromosomes; m0 represents the female risk chromosome and M1 represents the female normal chromosome. The detection result indicates that the embryos of P917-JJ-2, P917-JJJ-3, P917-JJ-5, P917-JJ-6 and P917-JJ-7 are not inherited to risk chromosomes of the female.
TABLE 2 test results
The quality control results are shown in Table 3, and it can be seen that the average sequencing depth is above 100 × and the coverage of 30 × is above 70%, and the coverage of 100 × is above 50%, and the quality control is qualified.
TABLE 3 high throughput sequencing quality control
| Numbering | Name (I) | Average sequencing depth (X) of target region | Coverage degree | 30X coverage | Whether it is qualified or not |
| K05053 | Female prescription | 600.28 | 99.25 | 90.00 | Qualified |
| K05226 | Male prescription | 602.99 | 99.01 | 90.04 | Qualified |
| K05054 | Father of female | 512.53 | 99.06 | 87.03 | Qualified |
| P917-JJJ-1 | JJJ-1 | 433.22 | 99.58 | 89.49 | Qualified |
| P917-JJJ-2 | JJJ-2 | 479.66 | 99.41 | 89.63 | Qualified |
| P917-JJJ-3 | JJJ-3 | 487.35 | 99.40 | 90.71 | Qualified |
| P917-JJJ-4 | JJJ-4 | 500.65 | 99.29 | 86.45 | Qualified |
| P917-JJJ-5 | JJJ-5 | 578.49 | 99.16 | 90.07 | Qualified |
| P917-JJJ-6 | JJJ-6 | 647.77 | 99.34 | 89.83 | Qualified |
| P917-JJJ-7 | JJJ-7 | 881.98 | 99.53 | 92.48 | Qualified |
The results of the mutation detection are shown in Table 4, wherein Hom indicates that the mutation site is homozygous, Het indicates that the mutation site is heterozygous, Hemi indicates that the mutation site is hemizygous, and Normal indicates that no mutation is detected at the mutation site. Since the design scheme covered the coding region of BRCA1 gene, gene mutations occurring in the coding region were efficiently detected and the sequencing depth was greater than 100X.
TABLE 4 high throughput sequencing mutation detection
The effective SNP site statistics are shown in Table 5, and it can be seen that within 1M of both sides of the BRCA1 gene of male and female, there are more than 2 heterozygous SNP sites, which meets the requirements of the European human reproduction and Embryo Society (ESHRE) "for PGT practice guideline based on amplification" (2011 edition).
TABLE 5 statistics of SNP sites of target genes
SNP haplotypes (results of SNP of selected embryo parts) are shown in Table 6, wherein F0 and F1 represent normal chromosomes in men; m0 represents the female risk chromosome and M1 represents the female normal chromosome.
TABLE 6 SNP haplotypes
Example 2
24 carriers of BRCA1 were detected by the same detection method, and 22 exons of BRCA1 pathogenic gene were covered by the mutation site. The standard setting is that more than 3 effective SNPs are respectively arranged at two sides of the mutation site, and the passing rate of 24 examinees is 100%, which shows that the detection method of the invention has higher universality and accuracy.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
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<213> Artificial Sequence (Artificial Sequence)
<400>28
tgcttctcaa gggagtatgc 20
<210>29
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>29
aatacgggga gtgttttggt 20
<210>30
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>30
gaaagtcatc tcctccaggg 20
<210>31
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>31
tttgcagaaa ggcaggaaac 20
<210>32
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>32
tcagtccctc tgctctcag 19
<210>33
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>33
aagacagatt cagcaggagc 20
<210>34
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>34
tgtgagagga tgagaggagt 20
<210>35
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>35
ccacgcacct ctccataaaa 20
<210>36
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>36
gagatcccag ttcccttctg 20
<210>37
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>37
ggatgttctc caccgacg 18
<210>38
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>38
cacttactcc ccagtgacg 19
<210>39
<211>17
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>39
ctgtcctcgc ctgacac 17
<210>40
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>40
aagaggtagc ccagaggac 19
<210>41
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>41
agcttgctag tgcttcagag 20
<210>42
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>42
gggggttgtg agataaggag 20
<210>43
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>43
aagggaggta accattctgc 20
<210>44
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>44
tttccctgat taagccctgg 20
<210>45
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>45
agacaatagg gtcctcccag aa 22
<210>46
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>46
gggcagagtt tatgttgtga ttgg 24
<210>47
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>47
gccaaaaaca caaccttcct gt 22
<210>48
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>48
cgaagagaca gaaagtgtgt gtgt 24
<210>49
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>49
accgtgaggt gatgagattc 20
<210>50
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>50
acctctttag gagaagcagc 20
<210>51
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>51
ccatagtctt ctgcaaggga 20
<210>52
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>52
aaagttgact atccccaggc 20
<210>53
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>53
atcacccacc tctttcccta 20
<210>54
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>54
aaagttgact atccccaggc 20
<210>55
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>55
tcagtacaga gacctccctt 20
<210>56
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>56
ctgagttaag tgggactgcc 20
<210>57
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>57
agattctcca catccaccac 20
<210>58
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>58
gtgtgtatct tgcttggctg 20
<210>59
<211>15
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>59
ctctgcgctg ggagc 15
<210>60
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>60
catggtcatt gtcctcctga 20
<210>61
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>61
ggggtggagt ttggattttt 20
<210>62
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>62
cctaggcttt ccctctcttg 20
<210>63
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>63
tcttaaacct agacttgggg g 21
<210>64
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>64
accaatctca gttaaggaag gt 22
<210>65
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>65
ccacaggaag atgacccttt 20
<210>66
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>66
ttgtggtgtg gaagtctagg 20
<210>67
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>67
ggcactgttg cagatgaaaa 20
<210>68
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>68
tttaagccct caacttcccc 20
<210>69
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>69
tcctcaaggg acactactga 20
<210>70
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>70
gatggaacta gatgtgggca 20
<210>71
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>71
ccctgcagct actacttgag tc 22
<210>72
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>72
actgagatac agactgggat actgag 26
<210>73
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>73
ctatgagcat cgtgcagatg ga 22
<210>74
<211>28
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>74
tgtgtataaa tacttgccga aggtcaag 28
<210>75
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>75
ctcctgcatc cagagtcac 19
<210>76
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>76
ttctggatgt tttcttgggc 20
<210>77
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>77
gtcgggagga aaagcaaacg tc 22
<210>78
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>78
ccatggtgga agcaaatgat tcc 23
<210>79
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>79
cccgatcaat aattatcctg ggtttg 26
<210>80
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>80
ctgtttattc tttcttctct agagagagca 30
<210>81
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>81
ctgggacgtg ttgagttcta 20
<210>82
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>82
tcagaaggag tcatttgcca 20
<210>83
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>83
accctgtttg gaatactcta gtgga 25
<210>84
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>84
ctcctcaaat gtctggttga tctga 25
<210>85
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>85
tgaactgaat gaagacgcca ttaca 25
<210>86
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>86
tgcttacaac cttgactccc ttt 23
<210>87
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>87
cccaatccct taaaacaatg aactgc 26
<210>88
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>88
acctcattgg tttccttgtc cttg 24
<210>89
<211>27
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>89
ggaagtaaag cttttaaaag ccacgta 27
<210>90
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>90
cctgctgaag ggaatggaaa gaa 23
<210>91
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>91
ggaaccggag gttattggat 20
<210>92
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>92
tcacctgctt catttcctgt 20
<210>93
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>93
ggaaaaccag agagagcttc tgt 23
<210>94
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>94
ggaaaacatt cacatctgga tttaactgaa 30
<210>95
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>95
ggacttccct gtgagacatt 20
<210>96
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>96
gtttccaaac ggggaacttt 20
<210>97
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>97
tagggccaga aaagcttagg 20
<210>98
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>98
tatgcagttc tgtctgctgt 20
<210>99
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>99
taacaagcaa acagaggtgc 20
<210>100
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>100
ccgagcagtg tgtttatgc 19
<210>101
<211>17
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>101
ctcctggtgc tgacgac 17
<210>102
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>102
agcatagagg tgcgtgatg 19
<210>103
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>103
cctttcctct ttaccccagg 20
<210>104
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>104
ttacatatgg aggccgaagc 20
<210>105
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>105
ccaacacagg taggcagtaa 20
<210>106
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>106
gagaaggcag atggacagaa 20
<210>107
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>107
catcaaggtg aacagggagg 20
<210>108
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>108
ctcacaaact caatggggtg 20
<210>109
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>109
aaataaaacc acccgttttc agatgg 26
<210>110
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>110
gctggctaca gagagagatt gaaga 25
<210>111
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>111
gacgccaagg aacgatagg 19
<210>112
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>112
atttatttca ggctctgtcc ca 22
<210>113
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>113
gtggaaaagt ttgttagccc a 21
<210>114
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>114
tagcacctta ctccttcact g 21
<210>115
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>115
acaccattca gcatcagtct 20
<210>116
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>116
gggaaggaag gagagcaaaa 20
<210>117
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>117
ctgtcactgt ggttgtccat 20
<210>118
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>118
ggcttagagc cagattcaga 20
<210>119
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>119
ttacctatca atcctcaccc c 21
<210>120
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>120
ggctagactt cagtgtccgt 20
<210>121
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>121
ccttaagtat cttactagat tggtg 25
<210>122
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>122
gtgtgtttgt gttcacgata 20
<210>123
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>123
ctcttccaca tttctaagcg aa 22
<210>124
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>124
ccccagaaac ttggatagtc t 21
<210>125
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>125
atcagttgcc agtgtcttca 20
<210>126
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>126
gaaggaaagg aggactgagg 20
<210>127
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>127
agaaaggtac taggtcccca 20
<210>128
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>128
gtaggtgatc ctcttgagcc 20
<210>129
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>129
ctcaagaagc ccagaggatt 20
<210>130
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>130
gtctgtcagt ctgtccctg 19
<210>131
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>131
tggataaccc agaccgaga 19
<210>132
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>132
tttccagaac tcgaacccaa 20
<210>133
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>133
gtaactgcca gcttccaatg 20
<210>134
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>134
ggaaccccat aacgcagag 19
<210>135
<211>17
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>135
gtatgcacgc tccctgg 17
<210>136
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>136
agctttaacc ggctatccc 19
<210>137
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>137
gatgttttca ggaaggggga 20
<210>138
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>138
caagggaagg agccttactg 20
<210>139
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>139
ctggtgttcg attcaacaac gtg 23
<210>140
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>140
ggtagtcttg ggaggtcagc tt 22
<210>141
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>141
aatgaataca aaccacggcg 20
<210>142
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>142
ctgcctctta cggtttggta 20
<210>143
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>143
aggtctttga acactaggag g 21
<210>144
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>144
ctagaaaagt ctgtccgtgt g 21
<210>145
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>145
acgaaggtca ccagaaactg 20
<210>146
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>146
ttgcgatcag aggacaaagt 20
<210>147
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>147
cgcttctcct ccctggtttt g 21
<210>148
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>148
cggaaccggg atctatttcg g 21
<210>149
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>149
gcatcgttaa cccagattcc ctt 23
<210>150
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>150
gccagggcaa agatggtaat ga 22
<210>151
<211>28
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>151
gtttacctgt tatggatgaa actgacct 28
<210>152
<211>29
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>152
ctgtaactaa taaaacgtag gcacagaga 29
<210>153
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>153
gtgtgtttat ggggtttggg 20
<210>154
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>154
gatggagcag aatgagacca 20
<210>155
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>155
tgatcttcca gcagacagtg 20
<210>156
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>156
ttgcagcaaa aagtcttcgt 20
<210>157
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>157
tcaatatctt gcactctggg ttc 23
<210>158
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>158
aggaaacgag tagattggca c 21
<210>159
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>159
caactccagg tcagttaact aagtcc 26
<210>160
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>160
ccttggagtc ctgagtggaa ac 22
<210>161
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>161
tctgcagaaa tgggcaagtt cat 23
<210>162
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>162
ctgaaaggca gtgggagcac at 22
<210>163
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>163
ctgtggatat ccccttggac 20
<210>164
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>164
aggacaccaa tgaggaaagg 20
<210>165
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>165
gctgctggac aagctagaga tc 22
<210>166
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>166
cttacctcag ccttctggat ctc 23
<210>167
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>167
tcccaggtct ctcaaaaagc 20
<210>168
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>168
ccacttcaga aaccccagt 19
<210>169
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>169
actggggttt ctgaagtgg 19
<210>170
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>170
gggtcatccg atctttgtct 20
<210>171
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>171
gcggttggtt ctttctcttc 20
<210>172
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>172
ggtgacatca tagctttccg 20
<210>173
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>173
ggagagagta tgggaaaggc 20
<210>174
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>174
atgctaggct tggtttgaga 20
<210>175
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>175
caaggatcga gccaaaagc 19
<210>176
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>176
gaaacgatga tacccttctg c 21
<210>177
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>177
tattcctgaa aaggcaggct 20
<210>178
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>178
tgagttgatg agtctcggtg 20
<210>179
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>179
cattaatttg ctaaattgct ggctaagaca 30
<210>180
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>180
ggctgatggg aaagagcaac at 22
<210>181
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>181
gggaatggag agaaggaaaa tctagt 26
<210>182
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>182
gcctcttatt aaacatacag aaggaccttt 30
<210>183
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>183
gcctcgcctc atgtggtttt at 22
<210>184
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>184
aaatagttcc aggacacgtg tagaac 26
<210>185
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>185
cggccatgca attattttta ttatgaagtg 30
<210>186
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>186
cccttgtctc acatgggtga at 22
<210>187
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>187
gttgctcctc cacatcaaca ac 22
<210>188
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>188
cctgctttta aacagctggg agat 24
<210>189
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>189
gcagagggaa ggctcagata ca 22
<210>190
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>190
cttaacttgt ttacagcgat gccaa 25
<210>191
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>191
atgttggagc taggtcctta ctct 24
<210>192
<211>28
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>192
actaggtgat ttcaattcct gtgctaaa 28
<210>193
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>193
ccctaatcta agcatagcat tcaattttgg 30
<210>194
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>194
ggactcatta ctccaaataa acatggactt 30
<210>195
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>195
attcctcttc tgcatttcct ggatt 25
<210>196
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>196
ggtactgatt atggcactca ggaaa 25
<210>197
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>197
tggttctgtt tttgccttcc 20
<210>198
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>198
aatgctgaag accccaaaga 20
<210>199
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>199
cttccctaga gtgctaactt ccag 24
<210>200
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>200
atggaaggta aagaacctgc aact 24
<210>201
<211>27
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>201
cttgggatat tcaacactta cactcca 27
<210>202
<211>27
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>202
gcaatgcatt atatctgctg tggattt 27
<210>203
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>203
gcgcttgtac ttgtcaacag tt 22
<210>204
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>204
agaagattgg ctcttaccac ttgtc25
<210>205
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>205
gtggagggta ctttcccag 19
<210>206
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>206
ttctggctga aggtggaag 19
<210>207
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>207
agccttgaag gagatgagtg 20
<210>208
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>208
tggatggaac taacaaggac a 21
<210>209
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>209
ttttagagtg acattgcact gatg 24
<210>210
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>210
tgttgaactg ggaggcatat ta 22
<210>211
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>211
tttacctccc tggctttctg 20
<210>212
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>212
gtccgttcag gggaacataa 20
<210>213
<211>27
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>213
ccctccttgt tttattgact cttcagt 27
<210>214
<211>27
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>214
cacatttctg atccagatca catgtct 27
<210>215
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>215
taattggcac cgttgctttc 20
<210>216
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>216
tgctcaatag gtgctggtag 20
<210>217
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>217
tgtaggcaga ttggtctctt tt 22
<210>218
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>218
aagcttgtta gcactgacgc 20
<210>219
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>219
ccgtgggtgt aatatggtga 20
<210>220
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>220
aaagaacgca cacactaagc 20
<210>221
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>221
aacccggatt taagcgtgta 20
<210>222
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>222
cgaggtgtag ttttcgaagc 20
<210>223
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>223
aacccggatt taagcgtgta 20
<210>224
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>224
cgaggtgtag ttttcgaagc 20
<210>225
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>225
gtaaacgcaa cacacaacct 20
<210>226
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>226
ccttgatcgc tggaaggatt 20
<210>227
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>227
cgttgacaat ctcccgttag 20
<210>228
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>228
gtggccgatc gttaacattt 20
<210>229
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>229
aaatggccca atgcaaacac 20
<210>230
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>230
agtacagtgg ccgatcgtta 20
<210>231
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>231
tgcaaacacg ttgacaatct 20
<210>232
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>232
gtacagtggc cgatcgtta 19
<210>233
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>233
tgcaaacacg ttgacaatct 20
<210>234
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>234
gtacagtggc cgatcgtta 19
<210>235
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>235
ggatagtgaa acacatgcgg gaaa 24
<210>236
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>236
caaggtggaa tgggagcttc tt 22
<210>237
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>237
gcacgtttga caatcctgtg tct 23
<210>238
<211>28
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>238
atctcctgtg ctacatgcaa atatacaa 28
<210>239
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>239
ctgggcaaca caccactgga at 22
<210>240
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>240
cgggtagttt aggatagttg gtaggt 26
<210>241
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>241
gccctccaag taaagtcgaa 20
<210>242
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>242
tctcctggga tacaacctgt 20
<210>243
<211>29
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>243
acaacaacac aaaaaccaag agaaatctt 29
<210>244
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>244
ccactggtga cgacgtaaag at 22
<210>245
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>245
gggatgcgat ggtagtgaga at 22
<210>246
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>246
gcctagggtt cggccttaaa aa 22
<210>247
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>247
aagcagcata gcagcagaga tt 22
<210>248
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>248
gctctaattg aggcctgtca caa 23
<210>249
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>249
ccagtctctg ctggctcttt ag 22
<210>250
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>250
gactttctat aaatccggtt ttctcatcca 30
<210>251
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>251
caagcctcag gcttgtatgt ttg 23
<210>252
<211>28
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>252
gaggataatt aactctggaa catcaggt 28
<210>253
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>253
aattatgagg ggccacagag 20
<210>254
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>254
cagaactgtc tctgtccctg 20
<210>255
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>255
cagttatgac aacaggccat gaac 24
<210>256
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>256
gcccagatgc agagaggtta ga 22
<210>257
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>257
ggcttcaggg taaagctatg 20
<210>258
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>258
tctgtgagac aaactcagca 20
<210>259
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>259
cggaactggc ttctctcttt 20
<210>260
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>260
acctagtttc cagaaccacc 20
<210>261
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>261
caggggagca gtaacaactt 20
<210>262
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>262
ctaccatacc agtggacacc 20
<210>263
<211>29
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>263
aaacaataat tgcggaaagt gatgaaagt 29
<210>264
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>264
tggagggaag tagcctgaag tt 22
<210>265
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>265
gacagataga gaccttcagt gc 22
<210>266
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>266
ggacctaagc ctagggagat a 21
<210>267
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>267
cctctaggct cctggacaca ta 22
<210>268
<211>27
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>268
tgagagttct aggattctct ggtgatg 27
<210>269
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>269
atcccagaaa cagagagcag 20
<210>270
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>270
ctgagcctgc ctgtattcaa 20
<210>271
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>271
tgacctggga atctgcatct ttt 23
<210>272
<211>28
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>272
aacctattta ccagaaatgg ctttgttc 28
<210>273
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>273
tgactagcag atactgggga 20
<210>274
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>274
tggcttgcag ggatacatag 20
<210>275
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>275
accttctgac ctcaaagtgg 20
<210>276
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>276
ctattgtgct ctgatgctgc 20
<210>277
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>277
ctgacctcaa agtggctctg at 22
<210>278
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>278
actggctatt gtgacctcca aag 23
<210>279
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>279
aggccaatac aagaggtagc 20
<210>280
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>280
gccaacttct ttacaagggc 20
<210>281
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>281
cttccccacc cctagaaatc 20
<210>282
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>282
acctggctaa gaacatggag 20
<210>283
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>283
tctagattgt aagccccgtg 20
<210>284
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>284
ggtccatatg tgattaacgc tg 22
<210>285
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>285
gatctgttct gggactcctg tt 22
<210>286
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>286
agtgaggcag ggatctgagt ac 22
<210>287
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>287
agggctatcc cagcgtta 18
<210>288
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>288
ctttaggcct tggttaggag a 21
<210>289
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>289
cagtctttgc ttgggctatg 20
<210>290
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>290
taatttgctc tgtggagggg 20
<210>291
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>291
gctgaggaca gatgtcccta ct 22
<210>292
<211>26
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>292
agtaacttgg tagtgagaag agctga 26
<210>293
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>293
gggcaggatt ctggttacaa 20
<210>294
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>294
ttctcacaca cacagactgg 20
<210>295
<211>30
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>295
aaagaaatgg attacattct ggaaacttgc 30
<210>296
<211>24
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>296
ctgtcaaagc tatctgggtt cgat 24
<210>297
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>297
ccaggtctac gacactcaag 20
<210>298
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>298
gtttgcaacc ttaggagcac 20
<210>299
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>299
ttccccaaca cagagaagac 20
<210>300
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>300
tgacaagagt gttggaggag 20
<210>301
<211>25
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>301
cctaacctcc tagagttcac agtct 25
<210>302
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>302
ggctgaacac caccatagga at 22
<210>303
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>303
ggagactagg accacatcag gt 22
<210>304
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>304
ggactcccag gtggtgagaa ta 22
<210>305
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>305
cctgtctcta cttccagcag 20
<210>306
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>306
tgcttgccag gactttatca 20
<210>307
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>307
gtctccttac ctgtttggct 20
<210>308
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>308
tccctatcca ccctgaagaa 20
<210>309
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>309
ctgggtgttt tccctgagag 20
<210>310
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>310
accaggccca atatcctcta 20
<210>311
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>311
gtccttcagg agtgcagatt 20
<210>312
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>312
tagggtaggc atcctcatca 20
<210>313
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>313
tggccacttc cttaaacact 20
<210>314
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>314
tcacttggaa tccctcccta 20
<210>315
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>315
ctcaggctgt aagcaagaga 20
<210>316
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>316
ttcagtcaca gacactgacc 20
<210>317
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>317
gagactgtct gccttccttt 20
<210>318
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>318
attaaattgg ggcttggtgc 20
<210>319
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>319
atctgcagct gctgtgtg 18
<210>320
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>320
tccctagact tccaacgaga 20
<210>321
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>321
ttttccagga agaggtgagg 20
<210>322
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>322
gtgccaatct ttctgaccac 20
<210>323
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>323
cactccccct cttcatcatc 20
<210>324
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>324
tttgtgagta cagtgggctt 20
<210>325
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>325
ctattaccac ctcccacacc 20
<210>326
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>326
gtctgtcgta attgttgccc 20
<210>327
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>327
agaatgttag gcctgggaag 20
<210>328
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>328
gtggttagct tgaggcaatc 20
<210>329
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>329
acatgtatga cgggagcctg 20
<210>330
<211>23
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>330
atgtaaattg aatgtggctg tca 23
<210>331
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>331
tggacggcct attatcatca 20
<210>332
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>332
gaaaaggttg ccttcgctat 20
<210>333
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>333
cctaggaaag gctgagttgt 20
<210>334
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>334
acagaagcta aggttcaggg 20
<210>335
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>335
gaatggggta taggagcctg 20
<210>336
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>336
gcaagcccat tttctctcag 20
<210>337
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>337
atcccactcg ggagagttc 19
<210>338
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>338
ctcaaggggg atgtgacc 18
Claims (10)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010338499 | 2020-04-26 | ||
| CN2020103384992 | 2020-04-26 |
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| Publication Number | Publication Date |
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| CN111575378A true CN111575378A (en) | 2020-08-25 |
| CN111575378B CN111575378B (en) | 2023-06-13 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202010441563.XA Active CN111575378B (en) | 2020-04-26 | 2020-05-22 | Detection method, detection composition and detection kit for hereditary breast cancer and ovarian cancer syndrome |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113005194A (en) * | 2021-04-27 | 2021-06-22 | 北京嘉宝仁和医疗科技有限公司 | Primer composition, product and method for detecting abnormal haplotype of chromosome small segment |
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