CN113403380A - Complex disease related SNP site primer composition and application - Google Patents
Complex disease related SNP site primer composition and application Download PDFInfo
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Abstract
The invention provides a complex disease related SNP locus primer composition and application thereof, belonging to the technical field of molecular biology. According to the invention, a group of SNP locus amplification primers are designed and developed by an NGS method, and the gene structure change related to the complex disease in an individual body is detected by a bioinformatics analysis method, so that the individual disease risk is predicted, and the prevention and/or treatment effect of the complex disease is improved. The evaluation system and the detection method have the advantages of low cost, simple and convenient operation, good sensitivity, high accuracy and good repeatability, and have great clinical application value.
Description
Technical Field
The invention relates to the technical field of molecular biology, in particular to a complex disease related SNP site primer composition and application thereof.
Background
Complex Disease (Complex Disease) is a general name for a Disease with complicated etiology, hidden onset, long course of Disease and persistent Disease. In recent years, the incidence and mortality of chronic complex diseases have been on the rise with the progress of rapid township and aging of population. Epidemiological investigations have shown that the occurrence of chronic complex diseases is closely related to poor lifestyle and behavior. The monitoring of chronic complex diseases can find the health problems of the organism in time and provide correction measures aiming at the related problems. Early screening and early detection of chronic complex diseases are significant for reducing the harm of the chronic complex diseases, the prevention of the chronic complex diseases starts from the prevention and control of risk factors of the chronic complex diseases, and genetic background is one of the risk factors influencing the chronic complex diseases.
The complex diseases are caused by factors of complex regulation and control among multiple genes and environmental action, have obvious characteristics of genetic heterogeneity and phenotype complication, and are commonly seen as neoplastic diseases, cardiovascular diseases, autoimmune diseases, nervous system diseases and the like. Unlike the general monogenic diseases, which do not completely comply with Mendelian's law of inheritance, the development of complex diseases often involves multiple biological processes, such as the dysregulation of the network dynamics of interactions between multiple genes, proteins or related molecules, resulting in specific phenotypes and symptoms of various diseases. The early diagnosis of the complex disease is very difficult, the prognosis effect is poor, and the recurrence rate and the death rate are high, so the method has important practical significance for the research of the complex disease.
A Single Nucleotide Polymorphism (SNP) is a polymorphism in a DNA sequence caused by a variation of a single nucleotide (including four forms of transition, transversion, deletion and insertion), and generally, only two forms of transition and transversion occur, and the ratio of the two forms is about 1: 2. SNPs are widely present in the human genome, and occur in about every five hundred to one thousand bases, accounting for more than 90% of all known genetic polymorphisms, and are the most common heritable variations in humans. Single nucleotide polymorphisms occur most often in CG sequences, with C-allelic switching to T-allelic being most common, and this may be related to the ease with which cytosine is methylated to form thymine. The single nucleotide polymorphism is used as a third generation DNA genetic marker which is explored, has the advantages of large quantity, large density, wide distribution, strong stability, easy detection and the like compared with the first two generations of markers, and is beneficial to the application and popularization of the single nucleotide polymorphism in the medical field.
The patent CN108334749B discloses a method and a system for detecting the epistasis of a complex disease based on a chromatin regulation loop, wherein the chromatin regulation loop based on gene interaction is established by utilizing data such as chromatin remote interaction, SNP interaction which can influence the phenotype of the complex disease in the regulation loop is calculated according to the chromatin regulation loop, the SNP interaction related to the complex disease can be rapidly and accurately explored, and a potential target is provided for subsequent drug involvement and the like. Patent CN105354444B discloses a method for screening complex disease susceptibility SNP combination based on susceptibility SNP, which can quickly find out susceptibility SNP combination of specific disease from a large amount of data, can be applied to screening of any complex disease susceptibility SNP combination and prediction of disease genetic risk, and provides a new method for disease risk evaluation. However, the above prior art only provides an evaluation method or system for a certain complex disease, and does not provide a detection method or system for a plurality of complex diseases.
Therefore, it is highly desirable to construct a primer composition for related SNP sites capable of simultaneously targeting multiple complex diseases and use thereof, so as to improve the effect of preventing or treating chronic complex diseases.
Disclosure of Invention
Aiming at the defects, the invention provides a complex disease related SNP locus primer composition and application thereof. According to the invention, a group of multiple SNP sites related to the complex disease and amplification primers thereof are designed and developed by an NGS method, and the change of the gene structure related to the complex disease in an individual body is detected by a bioinformatics analysis method, so that the individual disease risk is predicted, and the prevention and/or treatment effect of the complex disease is improved. The evaluation system and the detection method have the advantages of low cost, simple and convenient operation, good sensitivity, high accuracy and good repeatability, and have great clinical application value.
In order to achieve the above object, the technical solution of the present invention is as follows:
in one aspect, the present invention provides a complex disease-associated SNP site primer composition, comprising: rs10229583, rs1048886, rs10811661, rs10814916, rs10886471, rs10906115, rs1111875, rs1116357, rs11257655, rs117601636, rs11787792, rs12000501, rs12010175, rs12549902, rs13266634, rs1359790, rs1421085, rs1436955, rs1470579, rs147538848, rs 0315505500, rs163171, rs 16316361329, rs 29955379, rs17584499, rs 177982, rs 18018095, rs1845900, rs187060802, rs1965305, rs 202773617640, rs 4120796, rs2206734, rs2237892, rs2237895, rs 223782237897, rs 22835637799, rs 70779, rs 70777943779, rs 7077563756375637563756375648, rs 70779, rs 7037563756375637563756375637563756375637569, rs 70729, rs 7037563756375637563756375637563756375637563756375637567, rs 703756375637563756375637563756375637563756375637567, rs 70729, rs 7037563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637567, rs1745, rs 70729, rs 707245, rs1745, rs 7245, rs 70729, rs 729, rs 70729, rs 707245, rs 7072563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756300, rs 729, rs1745, rs 703756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756300, rs1745, rs 729, rs1745, rs 7037563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756300, rs1745, rs 703756375637563756375637563756375637563756375637563756375637563756375637563756375637, rs10883795, rs11038167, rs11038172, rs1635, rs16887244, rs2142731, rs835784, rs10821944, rs 11900600673, rs12379034, rs 12617617617617656, rs13192471, rs1854853, rs2075876, rs2233434, rs2841277, rs2867461, rs3093024, rs3124, rs3781913, rs 65707075, rs10865331, rs13210693, rs17095830, rs30187, rs4552569, rs10036748, rs10845606, rs10847697, rs 8334, rs1167796, rs1234315, rs12494314, rs 128569, rs 13177851338574, rs 1388190, rs1913517, rs 22057596409, rs 2200926, rs 09575637567746, rs 31779, rs 317746, rs 31575648 5648, rs 704746, rs 445646, rs 445648 5646, rs 445648, rs 445646, rs 725648, rs 445648, rs 44575646, rs 725648, rs 445648, rs 445646, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 445648, rs 445634575648, rs 445648, rs 725634575648, rs 43545648, rs 4456345743545648, rs 445648, rs 725648, rs 445634575634575648, rs 445634575648, rs 445634575634575634575648, rs 445648, rs 445634579, rs 445648, rs 445634575648, rs 445634579, rs 445648, rs 445634575648, rs 445648.
Specifically, the primer compositions are shown in table 1 below.
TABLE 1 primer compositions
Specifically, the complex disease includes, but is not limited to, type 2 diabetes, hashimoto's thyroiditis, alzheimer's disease, allergic dermatitis, rheumatoid arthritis, ankylosing spondylitis, systemic lupus erythematosus, asthma, or schizophrenia.
On the other hand, the invention provides the application of the SNP locus primer composition in preparing products for detecting and/or evaluating complex diseases.
In yet another aspect, the present invention provides a product for detecting and/or evaluating a complex disease, which comprises the above-described SNP site primer composition.
Specifically, the product is an independent reagent or a kit.
In another aspect, the invention provides an application of the SNP site primer composition or the product in detecting SNP sites related to complex diseases.
In still another aspect, the present invention provides a method for detecting a complex disease-associated SNP site, which is a non-disease diagnosis and treatment method, comprising detecting a SNP site in a test sample genome using the above primer composition or product.
Specifically, the method comprises the following steps:
(1) extracting genome DNA;
(2) 1, performing multiplex PCR reaction;
(3) magnetic bead purification of round 1;
(4) performing multiplex PCR reaction of 2 nd round;
(5) magnetic bead purification for round 2;
(6) quantifying the library;
(7) detecting the quality of the library;
(8) performing computer sequencing on the library to obtain a fastaq sequence file, and performing quality evaluation, comparison and SNP (single nucleotide polymorphism) calling to obtain the genotype information of the SNP of the capture region of the sample to be detected;
(9) assigning the genotype of each SNP to the OR value of the disease in the population;
(10) multiplying all OR values of SNPs involved in a disease in an individual by the absolute genetic risk of the individual for the disease;
(11) the absolute genetic risk is ranked in a thousand human genome/Chinese human genome queue, namely the relative genetic risk of the individual aiming at the disease is ranked within 5% as low risk, above 95% as high risk and between 5 and 95% as normal range.
Further specifically, the quality evaluation software in the step (8) is FASTQC software, the alignment software is BWA software, and the SNP calling software is GATK software.
Compared with the prior art, the invention has the advantages that:
1. the invention provides an SNP locus related to complex diseases and a primer composition thereof, which can simultaneously detect and evaluate the risks of the complex diseases such as type 2 diabetes, hashimoto's thyroiditis, Alzheimer's disease, allergic dermatitis, rheumatoid arthritis, ankylosing spondylitis, systemic lupus erythematosus, asthma, schizophrenia and the like, thereby guiding people to prevent in advance from the aspects of diet, exercise, living habits and the like and prompting the long-term attention point of routine physical examination in hospitals in the future, and further improving the prevention and treatment effect of the chronic complex diseases.
2. The SNP locus primer composition and the detection method provided by the invention can obtain enough genetic information from trace peripheral blood for genetic risk assessment, have good sensitivity, simultaneously, the efficient and specific amplification primer can accurately obtain the SNP locus information, avoid non-specific amplification, greatly improve the accuracy, reduce the experiment cost and have simple and convenient operation. The acquired SNP combination is subjected to calculation and prediction of the possible future disease risk of the individual by a specific analysis method, and the method has great clinical application value.
3. The SNP locus information contained in the invention covers various diseases, N genotypes of susceptible loci of the diseases are obtained for each disease, the genotype allele of each susceptible locus has an own OR value, the OR value corresponding to the genotype of the locus covered by a certain disease is obtained according to the genotype information of the locus obtained by a sequencing result, and the overall genetic risk value of the disease is further calculated. For risk assessment of a certain disease of an individual, the absolute genetic risk of the individual is put into a sufficiently large population for comparison, so that the risk of the disease of the individual can be represented more accurately. Therefore, the invention determines the genetic risk of the detected individual by taking the established natural population of China and the genome population of thousands of people as references. If 95% of the individuals are cut-off and the OR value of a certain disease exceeds 95% of the people, the genetic risk of the disease of the individual is considered to be higher.
Drawings
FIG. 1 is a graph of in-flight sequencing data.
FIG. 2 is a diagram showing the genetic risk assessment of Alzheimer's disease.
Fig. 3 is a diagram of the genetic risk assessment of adult asthma.
FIG. 4 is a chart of the genetic risk assessment of Classification disorders.
FIG. 5 is a graph of genetic risk assessment of type 2 diabetes.
FIG. 6 is a graph of genetic risk assessment of Hashimoto's thyroiditis.
FIG. 7 is a diagram of the genetic risk assessment of systemic lupus erythematosus.
FIG. 8 is a diagram of the genetic risk assessment of ankylosing spondylitis.
FIG. 9 is a diagram showing the genetic risk assessment of rheumatoid arthritis.
Detailed Description
The present invention will be further illustrated in detail with reference to the following specific examples, which are not intended to limit the present invention but are merely illustrative thereof. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.
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.
In certain embodiments, details of the reagents or instruments used are as follows:
anhydrous ethanol: the Beijing chemical industry is analytically pure;
Agencourt AMPure XP Kit：Beckman(A63881)；
Qubit dsDNA HS Assay Kit：Life Technologies(Q32851)；
Nuclease-free water：Ambion(AM9930)；
High Sensitivity DNA Kit：Agilent(p/n 5067-4626)；
PCR instrument: ABI 9700 by Life corporation;
DynaMag-96 Side：Life Technologies(cat.no.12331D)；
Agilent 2100Bioanalyzer system：Agilent(G2939AA)。
example 1 primer composition
The primer compositions described herein are specifically shown in table 2 below.
TABLE 2 primer compositions
Example 2 library construction and detection
1. Genomic DNA extraction
Saliva or blood genomic DNA (gDNA) extraction was performed according to commercial genomic DNA extraction kit procedures and quantified using the Qubit (Life technologies).
2. 1 st round of multiplex PCR reaction
1.1. The reaction system is shown in table 3 below.
TABLE 3 reaction System
| Reagent | Volume(μL) |
| ddH2O | 4-x |
| Enhancer NB(1N) | 5 |
| Primer (Table 2) | 2.4 |
| gDNA | X(30ng) |
| IGT-EM101polymerase mixture | 3.6 |
Wherein the final concentration of each Primer in the Primer mixture is 0.5 pmol.
1.2. The reaction procedure is as follows: 30s at 95 deg.C for 3 min; 18 cycles at 98 ℃ for 20s and 60 ℃ for 8 min; 5min at 72 ℃.
3. Round 1 magnetic bead purification
3.1. Adding 15 mu L of AMPure XP magnetic beads which are balanced at room temperature into 15 mu L of PCR products, and sucking and uniformly mixing the products for a plurality of times by using a pipettor;
3.2. after incubation for 5min at room temperature, the PCR tube was placed on a DynaMag-96 Side magnetic frame for 3 min;
3.3. thoroughly removing the supernatant, taking the PCR tube off the magnetic frame, adding 40 mu LYF buffer B into the tube, and sucking and mixing the mixture for several times by using a pipettor;
3.4. after incubation for 5min at room temperature, the PCR tube was placed on a DynaMag-96 Side magnetic frame for 3 min;
3.5. removing the supernatant, continuously placing the PCR tube on a magnetic frame, adding 180 mu L of 80% ethanol solution into the PCR tube, and standing for 30 s;
3.6. removing the supernatant, continuously placing the PCR tube on a magnetic frame, adding 180 mu L of 80% ethanol solution into the PCR tube, standing for 30s, and completely removing the supernatant;
3.7. standing at room temperature for 10min to completely volatilize residual ethanol;
3.8. taking down the PCR tube from the magnetic frame, adding 22 mu L of nucleic-free water, gently sucking a pipette to beat the heavy suspension magnetic beads to avoid generating bubbles, and standing for 2min at room temperature;
3.9. placing the PCR tube on the magnetic frame again, and standing for 3 min;
3.10. pipette 9.4. mu.L of the supernatant, and transfer to a new 200. mu.L PCR tube, where the supernatant is the multiplex PCR product.
4. Multiplex PCR reaction round 2
4.1. The reaction system is shown in table 4 below.
TABLE 4 reaction System
| Reagent | Volume(μL) |
| Step 3.10 multiplex PCR reaction product mixture | 9.4 |
| IGT-I5(5μM) | 1 |
| IGT-I7(5μM) | 1 |
| IGT-EM101polymerase mixture | 3.6 |
4.2. The reaction procedure is as follows: 30s at 95 deg.C for 3 min; 20s at 98 ℃, 1min at 58 ℃, 30s at 72 ℃ and 8 cycles; 5min at 72 ℃.
5. Round 2 magnetic bead purification
5.1. Adding 13.5 mu L of AMPure XP magnetic beads which are balanced at room temperature into 15 mu L of PCR products, and sucking and uniformly mixing the mixture for a plurality of times by using a pipettor;
5.2. after incubation for 5min at room temperature, the PCR tube was placed on a DynaMag-96 Side magnetic frame for 3 min;
5.3. thoroughly removing the supernatant, taking down the PCR tube from the magnetic frame, adding 40 mu L YF buffer B into the PCR tube, and sucking and mixing the mixture by a pipettor for several times;
5.4. after incubation for 5min at room temperature, the PCR tube was placed on a DynaMag-96 Side magnetic frame for 3 min;
5.5. removing the supernatant, continuously placing the PCR tube on a magnetic frame, adding 180 mu L of 80% ethanol solution into the PCR tube, and standing for 30 s;
5.6. removing the supernatant, continuously placing the PCR tube on a magnetic frame, adding 180 mu L of 80% ethanol solution into the PCR tube, standing for 30s, and completely removing the supernatant;
5.7. standing at room temperature for 10min to completely volatilize residual ethanol;
5.8. taking down the PCR tube from the magnetic frame, adding 24 μ L of nucleic-free water or 1 × TE buffer (pH 8.0), gently pipetting the resuspended magnetic beads by a pipette to avoid generating bubbles, and standing at room temperature for 2 min;
5.9. placing the PCR tube on the magnetic frame again, and standing for 3 min;
5.10. pipette 20. mu.L of the supernatant, which is the prepared multiplex PCR library, into a new PCR tube.
6. Library quantification
2 μ L of library samples were used
3.0fluorometer (qubit dsDNA HS Assay kit) to determine the concentration of the library, record the library concentration.
7. Library quality detection
Using Agilent 2100Bioanalyzer system (High Sensitivity DNA Kit), 1 μ L of library sample was taken for library fragment length and purity measurement, the target fragment distribution interval of the normal library was between 300 and 400bp, and the main peak was around 360 bp.
8. And performing computer sequencing on the library to obtain a fastaq sequence file. And obtaining the genotype information of the SNP of the capture area of the sample to be detected through the steps of quality evaluation, comparison, SNP calling and the like, wherein FASTQC software is used for quality evaluation, BWA software is used for comparison, and GATK software is used for SNP calling.
9. The genotype of each SNP obtained is assigned to the OR value of the disease in the population study.
10. All OR values of SNPs involved in a disease in an individual are multiplied, i.e. the absolute genetic risk of the individual for the disease.
11. The absolute genetic risk is ranked in a thousand human genome/Chinese human genome queue, namely the relative genetic risk of the individual aiming at the disease is ranked within 5% as low risk, above 95% as high risk and between 5 and 95% as normal range.
Example 3
The correspondence between the complex disease and the gene and the detection site is shown in the following table 5.
TABLE 5
Experimental example 1 accuracy test
The 221 sites were tested using the assay method described herein and verified by Sanger sequencing, the results of which are shown in Table 6 below.
TABLE 6 accuracy test results
As can be seen from Table 6, the accuracy of the primer composition and the detection method described in the present application was 100%.
Experimental example 2 repeatability test
1 of the samples A and 1 of the samples B were selected, and each sample was tested in a batch by repeating 3 times, and the test results are shown in Table 7 below.
TABLE 7 results of repeated measurements
Remarking: sample A-1 is sample A at the 1 st test.
As shown in the test results in Table 7, the repeatability of the primer composition and the test method disclosed in the present application is 100%.
Experimental example 3 detection of precision
1 of the samples A and 1 of the samples B were selected, and each sample was tested with three batches of the primer composition, and the test results are shown in Table 8 below.
TABLE 8 results of precision measurements
As is clear from the results in Table 8, the precision of the primer composition and the detection method described in the present application was 100%.
Experimental example 4
1 sample of the peripheral blood A is selected to be detected according to the method of the application example 1 and example 2, and the genetic risk of the complex diseases is evaluated, wherein the genetic risk of the sample in the thousand human genome population and the current self-test genome population is shown in figures 2-9 by taking Alzheimer's disease, adult asthma, mental classification disease, type 2 diabetes, Hashimoto's thyroiditis, systemic lupus erythematosus, ankylosing spondylitis and rheumatoid arthritis as examples. Wherein, the abscissa is the natural logarithm of the normalized relative risk OR value, and the ordinate is the probability density of the crowd on the corresponding OR value. Alzheimer's disease OR ═ 1, RankThousand human genomes＝20％，RankSelf-test50 percent; adult asthma OR 1.1825, RankThousand human genomes＝3.7％，RankSelf-test13.6 percent; mental classification OR 1.4222 RankThousand human genomes＝85.6％，RankSelf-test94.97%; type 2 diabetes OR 0.4305, RankThousand human genomes＝7％，RankSelf-test23.6%; hashimoto's thyroiditis OR ═ 1, RankThousand human genomes＝0.1％，RankSelf-test0.2%; systemic lupus erythematosus OR 1.8085, RankThousand human genomes＝76.3％，RankSelf-test51.2 percent; ankylosing spondylitis OR ═ 1.26, RankThousand human genomes＝6％，RankSelf-test3.7 percent; rheumatoid arthritis OR ═ 1.11, RankThousand human genomes＝1.9％，RankSelf-test＝2％。
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 present 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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<400> 12
gcctggaagc ctctctccta 20
<210> 13
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 13
ttgcaaagaa gtgtagaagg aagaga 26
<210> 14
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 14
taacttctca ctcccttcca cct 23
<210> 15
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 15
gggagtttat taaattcctt gaagtagtcc 30
<210> 16
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 16
ctctaagggc aaaactaaga gttattgac 29
<210> 17
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 17
gctaggtaaa tatttgcaaa gcacct 26
<210> 18
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 18
gccactaaaa tgtcccaatg cc 22
<210> 19
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 19
attgctgggt cagatggtag ttc 23
<210> 20
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 20
cacaccagtc agaatggcaa ttattaa 27
<210> 21
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 21
gccacctgac acaagggg 18
<210> 22
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 22
cgcccaggaa aagcaggt 18
<210> 23
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 23
gtttaaagca aaaatgttaa tttagtgttt caca 34
<210> 24
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 24
ttcccactac actagatcct gaaaatataa a 31
<210> 25
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 25
ccaccagggt tcacaagaca a 21
<210> 26
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 26
atgtgtgttg tttaagtggc tgaatt 26
<210> 27
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 27
ctgcacgcat gttcttgcc 19
<210> 28
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 28
aagcatatgc atgttctgtg attcag 26
<210> 29
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 29
gattatcaga gcaaacgtgg cttc 24
<210> 30
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 30
tttctctccg aaccacttgg c 21
<210> 31
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 31
tgttgtcaac agtgagatat atcctcatta 30
<210> 32
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 32
caaatagcaa gttagctggg cataaa 26
<210> 33
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 33
ttttatagta gcagttcagg tcctaagg 28
<210> 34
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 34
atgcttctgg acagtgcgta g 21
<210> 35
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 35
atgtgcaagt aatctctgct ctacat 26
<210> 36
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 36
acactccttt taaatgatga agaaacaaaa c 31
<210> 37
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 37
gatgaaaaat ggctactgca actaaga 27
<210> 38
<211> 15
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 38
ggcccacggg ttgcc 15
<210> 39
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 39
ggattctgaa tgcactgaat atgtgtaata 30
<210> 40
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 40
gcgaggtggt gggcac 16
<210> 41
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 41
agaaagcagg gctggaaatc c 21
<210> 42
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 42
tctggctgct cagtagggg 19
<210> 43
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 43
cctctgcatt tttcccttgt gttt 24
<210> 44
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 44
ccggacctca gttctgcatc 20
<210> 45
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 45
ggaaatgtca attatgctgc tgaattaaaa 30
<210> 46
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 46
tgacacagtg gctgctcttt t 21
<210> 47
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 47
ggctattcca tatcgctgaa gagat 25
<210> 48
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 48
tcaaggtcta acatagcaaa tttcacatag 30
<210> 49
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 49
agctgattga atggaagact tctcata 27
<210> 50
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 50
tgactgatgg aggtgtggat tttt 24
<210> 51
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 51
agaatggaat agtaggccta agttacac 28
<210> 52
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 52
tcagcacatt tattttctgt gaaacca 27
<210> 53
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 53
attctttcct cccctcttaa tgcc 24
<210> 54
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 54
ggctagagat atcctataac cactccta 28
<210> 55
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 55
ttcctacttt gatgataagc actcagat 28
<210> 56
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 56
atgccatgtt ttaaatctct gcagg 25
<210> 57
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 57
taaagaagta ttgtgtcata gatggggaa 29
<210> 58
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 58
cacaatgtgc aggcttgtta catat 25
<210> 59
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 59
aaatctatct cacattccaa tccctgt 27
<210> 60
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 60
gtctggaaga aaatatatag ggtttgcttt 30
<210> 61
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 61
atgccagaag gacatattta ttgctttt 28
<210> 62
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 62
acttgagaca acatgagtga acca 24
<210> 63
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 63
taaccttaaa attggaaaca cttcttgcta 30
<210> 64
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 64
tgtttcagtg gaagttggca gata 24
<210> 65
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 65
atgttgtccc ttagcaccca tc 22
<210> 66
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 66
gttacttccc aactttgttg cttca 25
<210> 67
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 67
gtgtgtgtag aatgatattc cctcca 26
<210> 68
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 68
ggccgggacc tggagat 17
<210> 69
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 69
aactcagcat gggtaatagg tagttac 27
<210> 70
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 70
atttcaacat gaggtttgga gaatcaaa 28
<210> 71
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 71
gcatcctaag gtggttcaga gg 22
<210> 72
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 72
gtgtaaggca tctggtggag ag 22
<210> 73
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 73
gcagtgacag ggtgatcgg 19
<210> 74
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 74
catctgcctc ttggtctcat cttta 25
<210> 75
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 75
cgtcttgagc cagcaggg 18
<210> 76
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 76
gggaggggct cagggg 16
<210> 77
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 77
cgtcttgagc cagcaggg 18
<210> 78
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 78
gggaggggct cagggg 16
<210> 79
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 79
cacacaggag ccagcagg 18
<210> 80
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 80
atagacatgt gtttacccat cacgt 25
<210> 81
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 81
gtacttatac catgaaaaac cctgtacac 29
<210> 82
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 82
ctacgtgccc catcctgc 18
<210> 83
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 83
gtttcttaat gttatgtgat tagcaactaa aact 34
<210> 84
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 84
accactggag gtaaattact aattcatct 29
<210> 85
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 85
gtgggacctg ccaagcc 17
<210> 86
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 86
atgcagaaga tgggtccaac c 21
<210> 87
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 87
agagttaata tatgcaagac acttaggaca 30
<210> 88
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 88
aattctgatt ttgttttgtt ttgttctgtt ttt 33
<210> 89
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 89
gtccactcag ggttcctcct 20
<210> 90
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 90
accagtctgt ctgtccccc 19
<210> 91
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 91
ttcatttctt taaacatttg tttcgaatgg t 31
<210> 92
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 92
aagagtgcct ctagaacttg aaaagt 26
<210> 93
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 93
ttgcgtgtat atcttaatat ggataaagtt cc 32
<210> 94
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 94
tccagtaaga tttcgctgtc agag 24
<210> 95
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 95
gggaattctt aataggaatg actaaagcg 29
<210> 96
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 96
ctgggattag cgccaccag 19
<210> 97
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 97
ggtgagagaa ttacttgagc ccaa 24
<210> 98
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 98
caaaaacagt aaaaactaat acaccaggaa aata 34
<210> 99
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 99
gtacaattga gtgattttct gtatagtcac a 31
<210> 100
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 100
ccttgagaac cgtatgctaa gtga 24
<210> 101
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 101
tctccaaggt caatgaaaaa gaaaaatct 29
<210> 102
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 102
ttgcttgtct gaaaacgatt ttgtttc 27
<210> 103
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 103
ctcacactgc tataaagaca tacctga 27
<210> 104
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 104
actcccatct cctcctgcc 19
<210> 105
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 105
atagctgtac ttaaagagct tgtaggaat 29
<210> 106
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 106
ccaagccaca gaggagcg 18
<210> 107
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 107
aacaagagcc cagaaccaac tt 22
<210> 108
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 108
tatttagtgt gaagcaacaa caatcatcta t 31
<210> 109
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 109
caatgagacc ggtgtgggg 19
<210> 110
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 110
ccttagttct ccaaacaatt ctccatg 27
<210> 111
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 111
gccttttgat acaaaatacc atttggtg 28
<210> 112
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 112
gtgtgttatt atcacgtcta tgaatagcc 29
<210> 113
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 113
tttaatgaga tgtgactttt tatgacaatg aatg 34
<210> 114
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 114
aattaatata ccggtccata aaacatgctc 30
<210> 115
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 115
cacagagcag tggtctcagt c 21
<210> 116
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 116
gcgccgagga atctccg 17
<210> 117
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 117
cctattgcca ttctgaaaag ctgg 24
<210> 118
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 118
taaacatctt tgtaatatcc tgttatttac gct 33
<210> 119
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 119
tctgtctttt gagaagtttc cagtct 26
<210> 120
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 120
cccatacttg gtgactagct ctttaa 26
<210> 121
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 121
cccaaacata taagacttcc aatcctttat t 31
<210> 122
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 122
gaaatgaggc ttccaaacta gattgtag 28
<210> 123
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 123
cctctcttgc atgcgttttg g 21
<210> 124
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 124
ccaagaagta ctggaaggaa aacaaaa 27
<210> 125
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 125
taaagaaaga tgactctaag ggatttactg t 31
<210> 126
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 126
gatttctaag gtgaaaaggc aaccaa 26
<210> 127
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 127
tggatgggta agtagatgga tgga 24
<210> 128
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 128
tggaagattg ttgacatccc ctatc 25
<210> 129
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 129
gaccaaggca ggcgtcc 17
<210> 130
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 130
cttctcttaa tttcaattcc tttcattttc tagg 34
<210> 131
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 131
cccagctaac tttttgtatt tttagtgga 29
<210> 132
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 132
ccctgtggcc taggttcct 19
<210> 133
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 133
tggagtgttt cataaagtaa tcaatatcga c 31
<210> 134
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 134
cctgacctca gatgatctgc ac 22
<210> 135
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 135
gcgtgaactg gggaggc 17
<210> 136
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 136
tactaatatc ttaccccttt gtgtcacc 28
<210> 137
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 137
ccactgcccc atcctaccta 20
<210> 138
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 138
ggacgtttgg aggagcgg 18
<210> 139
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 139
ctcagctact tggaaggcta agg 23
<210> 140
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 140
tttgcatttt cctaatgatt agttatgttg atg 33
<210> 141
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 141
aagtgtgaat agagaaaatg ctcattacc 29
<210> 142
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 142
aaagcacaaa gtttttaata ttgatgagat cc 32
<210> 143
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 143
atgttgatta ttaggctgag tagtttgaac 30
<210> 144
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 144
tggcagtgaa tatacaggct gaatta 26
<210> 145
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 145
cagtgagcca agatcagcca t 21
<210> 146
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 146
agttgtatga tcatggttcg ctgta 25
<210> 147
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 147
tctggcatgc agctggc 17
<210> 148
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 148
taaaacacag tttgtaagca cccac 25
<210> 149
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 149
cttcaaagtt aacatttaat ttcttgtaat tgcc 34
<210> 150
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 150
cttcaaagtt aacatttaat ttcttgtaat tgcc 34
<210> 151
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 151
ttacattacc ttgtagtctg caggg 25
<210> 152
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 152
tagcaggctc ccactgtaca ta 22
<210> 153
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 153
aaaagggact attcctgtct acttcatag 29
<210> 154
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 154
tccccagaac ctcccctttt 20
<210> 155
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 155
cgtcatcacc cacccacg 18
<210> 156
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 156
atatctggtc actcctgctt tgtac 25
<210> 157
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 157
gctcgaagtc caaattcaaa atgttg 26
<210> 158
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 158
attagctggg caaggtggtg 20
<210> 159
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 159
ggagcaggga gtacagggt 19
<210> 160
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 160
tggggtttca ggtggggt 18
<210> 161
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 161
gctgtctttc ccagtagcta ctc 23
<210> 162
<211> 35
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 162
taaaaagaaa caaagaaatc tgtaacactg aatag 35
<210> 163
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 163
ataatggctg ggctacctcc t 21
<210> 164
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 164
atggtagaag atcttaattg ttagtgaaat gg 32
<210> 165
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 165
ccattgaatc taagctggac agaga 25
<210> 166
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 166
tcattttcca tattacagcc tttatgcaa 29
<210> 167
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 167
ggagggccca agagcaac 18
<210> 168
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 168
cgtgggtgaa atgtgacatt tagg 24
<210> 169
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 169
atcagttatg ccagtcccat taatttaaaa 30
<210> 170
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 170
agtcaacagt gttattttaa aactacttgg a 31
<210> 171
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 171
caggtagcag tgttcatatt cctga 25
<210> 172
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 172
cgtctaagtc ccttttcctc acatat 26
<210> 173
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 173
cgccactgca ctcaggag 18
<210> 174
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 174
gatctcagct caccatctgc c 21
<210> 175
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 175
accagcccgg ggagtac 17
<210> 176
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 176
tcggggctct gactggg 17
<210> 177
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 177
cattctgcct aatacagggt cttca 25
<210> 178
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 178
atagcaataa caacagaaga agagatgaac 30
<210> 179
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 179
ccctacatgc ctcttaccca ac 22
<210> 180
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 180
gttcagcaga gttggacagt ga 22
<210> 181
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 181
actttctgat tttctcaccc tacaca 26
<210> 182
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 182
gggaaatttc agcaaaagga ctgataa 27
<210> 183
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 183
atttcttttt cttgtcttgt attaactagg aca 33
<210> 184
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 184
tcctgtgaga atctaatgct gcc 23
<210> 185
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 185
ttctgaagtg attcaactat tttgagcata t 31
<210> 186
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 186
ttttcaccaa ggattcaaaa ataccctt 28
<210> 187
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 187
gtacaattga gtgattttct gtatagtcac a 31
<210> 188
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 188
ccttgagaac cgtatgctaa gtga 24
<210> 189
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 189
atgagatctc tgccggacca 20
<210> 190
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 190
actgtggttt ctcgtctgaa aacta 25
<210> 191
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 191
tggtaacgat ttctacatct gcagt 25
<210> 192
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 192
tctggcacgg agtcattctt g 21
<210> 193
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 193
aaccttacca attgaactaa ataagatacc ag 32
<210> 194
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 194
gactcttgtg cattcttcaa tatgtcat 28
<210> 195
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 195
atcttgggta aatatgtagg agtggaatt 29
<210> 196
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 196
aaaagacaat ccaaaagcca gcttta 26
<210> 197
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 197
caaggctatt ccatcctctt gaca 24
<210> 198
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 198
atgtgtttcc ttagtaactt ggggaa 26
<210> 199
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 199
ttaagtcctt gcttctttct ttgttcc 27
<210> 200
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 200
aaggttccag gggagttgga 20
<210> 201
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 201
ctgaaaccat ctcccaccca at 22
<210> 202
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 202
aaaatctata gctaacatca tacttaacgg tg 32
<210> 203
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 203
agatggggct aatgtttatt ggct 24
<210> 204
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 204
cttcatcaga gaccactcag agtaag 26
<210> 205
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 205
atctggatga aattacctaa aggagaaaat g 31
<210> 206
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 206
aggtattcaa ggggcgaaat cc 22
<210> 207
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 207
aggagaataa gggacagaca atgc 24
<210> 208
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 208
tgtcaccaca taatgcctct actg 24
<210> 209
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 209
cgtacagaca taaagtagat cagtggt 27
<210> 210
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 210
tcaagtgtac aattcagtgc tttttcata 29
<210> 211
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 211
tttttgtgga ggcggtcttc a 21
<210> 212
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 212
tgagaaacaa ctcaaacaag gatgtg 26
<210> 213
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 213
gggcttattc aaggtgacac aattag 26
<210> 214
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 214
cactctcact tccctctcat gc 22
<210> 215
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 215
ctgggcgcgg acatgg 16
<210> 216
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 216
gcggccctgt tccacc 16
<210> 217
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 217
agagttaggt cagcgaattc actc 24
<210> 218
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 218
gtagtaagac ttgagaggca ctgg 24
<210> 219
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 219
ttaatttgca tttccttgat ggttaatgat g 31
<210> 220
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 220
cagaatctgt aagtaacttt aacaattcaa acaa 34
<210> 221
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 221
ctgatcgtta ttaattcaga aagggtgtt 29
<210> 222
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 222
tttttaagga ggccaaaggg gaa 23
<210> 223
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 223
gagactggga tgtgatgagc ag 22
<210> 224
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 224
ctaaatgtct ccttcaatcc tcacctat 28
<210> 225
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 225
tgctaacagg actggtcaaa actaa 25
<210> 226
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 226
agttcctggc aaaacttgaa agaag 25
<210> 227
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 227
gcctgggttc accggc 16
<210> 228
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 228
gcggggtgaa aggtaggaag 20
<210> 229
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 229
gaatggaaaa tttgcctcat ttgcc 25
<210> 230
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 230
ctctggagca aatgatcaga acaac 25
<210> 231
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 231
gctgttatat agaagaggga ttatacttgg a 31
<210> 232
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 232
tacaatgtga gcagattaaa ctagttgatg 30
<210> 233
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 233
ctttggtaac catccttcta cttctctat 29
<210> 234
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 234
ccctgcgagg taggctct 18
<210> 235
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 235
gtgcagtcct gggctgtc 18
<210> 236
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 236
ggagtgtgtg tgctgttcca 20
<210> 237
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 237
gcttagcttt acctcttttg atattgaagt a 31
<210> 238
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 238
tgctacttac agattcactt cttatatggt 30
<210> 239
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 239
ctacataaga agttatctca aaatgtggca 30
<210> 240
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 240
gaagtgccac cttgagtata aatgtg 26
<210> 241
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 241
tttcgctttt cctagaatac attgcc 26
<210> 242
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 242
gccaaggctc tttgtcgtat ttac 24
<210> 243
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 243
ttcaatctcc atagtagtta taagtatgtt caga 34
<210> 244
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 244
atgaatcaga aatgaaagaa gagacgc 27
<210> 245
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 245
aactttggtg ctctccttta gttattttt 29
<210> 246
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 246
ccaaagggaa tcctgagagg c 21
<210> 247
<211> 35
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 247
ttgaggtata attacataaa ctgtatatta atgca 35
<210> 248
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 248
aacattatta tacattttga caacaacata tagc 34
<210> 249
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 249
gcctttcatc cctgagggc 19
<210> 250
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 250
ttgcctcaca taaaagtcat tgtgttaa 28
<210> 251
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 251
aaggtcacag tggcaaccag 20
<210> 252
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 252
aaccttccat caaattttgt tgactgta 28
<210> 253
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 253
atcagtgttt ccttagattt tattcaatac tcat 34
<210> 254
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 254
tgttgtgaaa atcagatgat ctaatgtctc 30
<210> 255
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 255
atctatttat tttcacagtt ggatgtactg g 31
<210> 256
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 256
tctgggttag aagtttgttt tctgaatg 28
<210> 257
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 257
taatgacagg ggcatgttct gag 23
<210> 258
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 258
tgcagtattc gatacagtaa cttcgt 26
<210> 259
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 259
attgatccca ggtctttgga ttataact 28
<210> 260
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 260
gccaaataag tgaccaatgg cc 22
<210> 261
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 261
ggcctctctt gaagtacatc atctt 25
<210> 262
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 262
ttgtagtcag ggatatagaa gaggaaga 28
<210> 263
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 263
ggcttcctag ataattaatt actgctgc 28
<210> 264
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 264
acctcgcctc catctccc 18
<210> 265
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 265
ggctgaagga gagagagagg at 22
<210> 266
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 266
gtttcttgaa cgactggtct tctttt 26
<210> 267
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 267
gttattagct ctttgtgatt ctaaagtgct a 31
<210> 268
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 268
gattacatgt ggcctacttc tttaaatctt 30
<210> 269
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 269
gttgactagg cggttctact gg 22
<210> 270
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 270
tggaatacag cagaaatgac attgtaag 28
<210> 271
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 271
gatgaggaac catgccagca 20
<210> 272
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 272
attcccagaa ttgctagcag atgt 24
<210> 273
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 273
aaagtgtata taaatcaact aaactgggca ata 33
<210> 274
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 274
tctccaaact ctctttaaag atacccatta ta 32
<210> 275
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 275
acattatggc tagagaagag taaagaagaa 30
<210> 276
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 276
tgctccatct cttctttctc tacaag 26
<210> 277
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 277
gaaaaaggca cctaaaataa aagagatgat g 31
<210> 278
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 278
ccgcattcta cacatgaggg aa 22
<210> 279
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 279
ttcatatttg agctgagcta ttatcatgc 29
<210> 280
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 280
atccttcagc catgcacaga g 21
<210> 281
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 281
tcttgctctg ttgcccaaac t 21
<210> 282
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 282
tggagtttgg tgccgaaaga a 21
<210> 283
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 283
atctgaaaaa taatttataa caggtgttct cct 33
<210> 284
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 284
attctctagt agggacaaaa acacatttta tag 33
<210> 285
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 285
atgccaacat cgaggtgtct c 21
<210> 286
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 286
gattagttaa gatgaggtca cactgga 27
<210> 287
<211> 15
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 287
cgctccccat ccgcg 15
<210> 288
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 288
gggcatgtcg gaggcg 16
<210> 289
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 289
ggacctgctt agtgttcgag ag 22
<210> 290
<211> 16
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 290
gggtgggcgg gctaga 16
<210> 291
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 291
ttataagtgt ctccatttta aagaacaggt tt 32
<210> 292
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 292
ttccaaggtg gcaaggctg 19
<210> 293
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 293
aggctcatgc tgtttgtccc 20
<210> 294
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 294
cgtgtgacgg atcagtgaag aa 22
<210> 295
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 295
gagcttgtct ctttttccct ttttca 26
<210> 296
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 296
aaaattgaag agacaaatgg gcagtt 26
<210> 297
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 297
gggatgagtt ggctgggc 18
<210> 298
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 298
tggcccctaa agcaaaactg t 21
<210> 299
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 299
ggaaacctag aagatgggca gag 23
<210> 300
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 300
catttcccta tcctccacag gc 22
<210> 301
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 301
aatggcatgc atgggaggg 19
<210> 302
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 302
agacaacagt caatttacaa cctaaagtac 30
<210> 303
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 303
ggaatattca ctgcatttga caggtt 26
<210> 304
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 304
atcttcaatc tcaccccaag cc 22
<210> 305
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 305
tgctacttca ccaatccgag c 21
<210> 306
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 306
agcaggccca ggcatcta 18
<210> 307
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 307
catgtagtgc tcttgcttca tgtg 24
<210> 308
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 308
gcataagctg gtgttctctc ttttc 25
<210> 309
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 309
actttggaac acttgaaatg agcaat 26
<210> 310
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 310
atatgaaagg cctccatttg tagct 25
<210> 311
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 311
atggttcatt gtggttttaa tttgctttt 29
<210> 312
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 312
tgtaacaaat ctgcacatat acctcct 27
<210> 313
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 313
aggaggtccc tgctcacc 18
<210> 314
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 314
tgtcagctcc tcagagggg 19
<210> 315
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 315
gggtgactac tttaagcctt actatctt 28
<210> 316
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 316
ccttagaact ctgtatagaa agctcaca 28
<210> 317
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 317
aaccgaagct aggatgttaa taaagaga 28
<210> 318
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 318
cagctttgtc actggttatg cg 22
<210> 319
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 319
ttttcatctt gcttctaaat tttcagcttt t 31
<210> 320
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 320
ttccaaagaa gtttcaagga accataaat 29
<210> 321
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 321
catgatgcca acctattcct tattcg 26
<210> 322
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 322
ccttagaggc ctttcatgtg aca 23
<210> 323
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 323
tttcaggcac agatttctga tagactat 28
<210> 324
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 324
agtaacgtat ttgacccctt taaaaacaa 29
<210> 325
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 325
gctgtagtga ggtctgaatg gg 22
<210> 326
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 326
catcgttttc cagacaaggc atc 23
<210> 327
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 327
cccaaagtgc tgagattacg ga 22
<210> 328
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 328
aaacttttct gaaaatctta agaggacaaa ac 32
<210> 329
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 329
ggctttcccc gtgttctcaa 20
<210> 330
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 330
tggcttcttc cttctcctgg a 21
<210> 331
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 331
atgtgaaaaa taccacgaca aaacca 26
<210> 332
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 332
gctgttaaca ggtacaaata aaatatctcc t 31
<210> 333
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 333
tatttttaga ttagacaggc tatcaatgga atc 33
<210> 334
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 334
accgcttggg ctccattg 18
<210> 335
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 335
aatcaggcaa tttggataga aaacagatta 30
<210> 336
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 336
gtgcttccat ttcaataatt ttatttggag t 31
<210> 337
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 337
ttggcagcaa gtaagggtgt t 21
<210> 338
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 338
tatgattgag acagacagtg ccg 23
<210> 339
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 339
ccttgtctta cttcttatta ttttgactac agat 34
<210> 340
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 340
tattgatgca actaactttc cctgattaag 30
<210> 341
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 341
ccacttctca gtacatgtgg gg 22
<210> 342
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 342
tatcatagca aagccccgtt tca 23
<210> 343
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 343
aatccttgtc acaagtcatg atcca 25
<210> 344
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 344
tggagtcagt tatgattctt tggca 25
<210> 345
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 345
aaggatggga aaagtctaca gataagg 27
<210> 346
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 346
gaaaacgatg actttgcttt tgtatcag 28
<210> 347
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 347
ggatgctgaa cctggaccag 20
<210> 348
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 348
aggaatccac agactcaatg ctc 23
<210> 349
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 349
gtaagaggga tgccacagtg g 21
<210> 350
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 350
cacaagccca ttctgtacta gatca 25
<210> 351
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 351
aatcatgaat aaagctgccg taaacat 27
<210> 352
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 352
ggctgaatca aaacagatcc aagg 24
<210> 353
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 353
ccagtccatt tgatcagcgg a 21
<210> 354
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 354
ctcgcacgtt tgacatcttt ctc 23
<210> 355
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 355
gtcttctaca cacacctctt ctgtt 25
<210> 356
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 356
tacaaactgg aagaggcatt gaca 24
<210> 357
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 357
aggaatagga gaaagctgcc atg 23
<210> 358
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 358
aaaagtaagt cctgtgggaa acatttc 27
<210> 359
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 359
gagctgagat ttaaagggag caga 24
<210> 360
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 360
gtaaactaat taagtgaaga tacaaagttg cag 33
<210> 361
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 361
gagccatgga ttgggaaagg t 21
<210> 362
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 362
tgaagtttct ggcacataga agtagtat 28
<210> 363
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 363
attttacaaa gcaaacaata tggcatagc 29
<210> 364
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 364
tgccagagta accctcctca a 21
<210> 365
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 365
atgataccgg cacaaagaga cc 22
<210> 366
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 366
tctgcatagt tatacagatt ccaagagtt 29
<210> 367
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 367
cctgtcattc tttgcccctg t 21
<210> 368
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 368
agcctgagaa cagacactcc t 21
<210> 369
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 369
tactatacat agaacaatgt ttgtcttccc ta 32
<210> 370
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 370
actcagagat aggatttact tgttggtag 29
<210> 371
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 371
cagtcatctt acaggccctt taaaac 26
<210> 372
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 372
atctcatgaa ggaaatgaac caaagaag 28
<210> 373
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 373
tcacaagaaa aattaaacac acatcatcaa g 31
<210> 374
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 374
tcttttcctg agcataataa tttagagatt catt 34
<210> 375
<211> 34
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 375
attcttaaat aaggacaaat tactcattag ctga 34
<210> 376
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 376
ggaggctttc ctaccccatt g 21
<210> 377
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 377
ctaatacagg ctaatatatt tgtggtgcc 29
<210> 378
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 378
tgttttctct gctcttggat agaataaatt att 33
<210> 379
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 379
gttctgagga tggtgcctgg 20
<210> 380
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 380
atgatgatgc tcacaactgc tct 23
<210> 381
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 381
ctgaaattgc cccagacagt ttc 23
<210> 382
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 382
atgcctggcc aaccacc 17
<210> 383
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 383
tcaatgctca agccccattt atttc 25
<210> 384
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 384
aattaagggg acacagtttt caaatgtaa 29
<210> 385
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 385
tgagggagct ttggaagaca ac 22
<210> 386
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 386
gcgcaggtgc tcactagg 18
<210> 387
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 387
ttacatgagt gtgtatgcag taaaagtatg 30
<210> 388
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 388
atttctttct gattgcttaa ataatgcaaa tct 33
<210> 389
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 389
gggaggtgtt gggtattcga aa 22
<210> 390
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 390
gttccactat ctgaccctcg ttc 23
<210> 391
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 391
gaggcatgga caatgaggca 20
<210> 392
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 392
atgatttgat aacatttccc actgaacat 29
<210> 393
<211> 33
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 393
aacaggatca catttataag tgtctaaagt aag 33
<210> 394
<211> 35
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 394
agaatataaa ttgactggta aataaggaag aaagt 35
<210> 395
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 395
attatctttg agtaggccct ttagaaacta 30
<210> 396
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 396
caacctactg agaattctcc aggg 24
<210> 397
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 397
aaaatatcag atgttgattc atgtaaggca 30
<210> 398
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 398
acccactcca cacccttttt c 21
<210> 399
<211> 35
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 399
ggagagatat atgtttaatg ttttagagaa atcca 35
<210> 400
<211> 35
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 400
atgctggtat taatatcttg aagatgaaga aatta 35
<210> 401
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 401
ggcctgggat tttaaaagtt ggaaa 25
<210> 402
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 402
ggttgcagca acttcaaaga gtt 23
<210> 403
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 403
ctcaaacact ccagatgaga aataagc 27
<210> 404
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 404
gcttctagct gtgaaagatg agttataaaa 30
<210> 405
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 405
tccttggcct cctcactagg 20
<210> 406
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 406
gacaatggtg actctggggt c 21
<210> 407
<211> 24
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 407
gcaagatcag ttggaggtat caga 24
<210> 408
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 408
tcctctctgt ccagttctca ca 22
<210> 409
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 409
gtaaccaaga gcttaaagac cattagc 27
<210> 410
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 410
ctgcggtcat cccccaac 18
<210> 411
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 411
tactttctgc cttcacatgt ccttatat 28
<210> 412
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 412
tgaagtcatg atggcaggcc 20
<210> 413
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 413
gtgagagaat gtgactgata aaaccaatt 29
<210> 414
<211> 35
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 414
aggttaacta ttatttttca cttacttttg agtct 35
<210> 415
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 415
ttggttcaga gaatcaaagc agaatc 26
<210> 416
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 416
gtttactttt tgctttctta atccatgct 29
<210> 417
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 417
caatatcata tggttataaa gggcagtgg 29
<210> 418
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 418
ctactttcct agtctttcag agctcaata 29
<210> 419
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 419
attgtagctg tatttagcag gaggaat 27
<210> 420
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 420
cctgctttca tttttcttag gtagatacc 29
<210> 421
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 421
gaagtctggt cccatgaata gattga 26
<210> 422
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 422
gattctccag gcttaaaact caatcag 27
<210> 423
<211> 32
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 423
aggattcctc ttctcttaga tataaaaact gt 32
<210> 424
<211> 28
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 424
caatttgcag ataacaagga agagatca 28
<210> 425
<211> 22
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 425
gcacgttttt ctctcagagc ac 22
<210> 426
<211> 17
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 426
tgggagcaga ggacggg 17
<210> 427
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 427
aacaactgct tgccctttag aaaat 25
<210> 428
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 428
acagcgattc ccctgaaaac a 21
<210> 429
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 429
ttggaacaga aggagggagg a 21
<210> 430
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 430
agacgtgcca gtttcctgac 20
<210> 431
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 431
attatgaaaa ctgtatccag agtgtgttta g 31
<210> 432
<211> 27
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 432
tcatgcaaga attcagattc agagttg 27
<210> 433
<211> 25
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 433
ttcaatcccc atatgcacag atctt 25
<210> 434
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 434
gatcaatcct tttacatatc ttctttggct 30
<210> 435
<211> 26
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 435
ccaagtagtg agagacatag aaccag 26
<210> 436
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 436
tgtgatattc tggtagagtt tccaatcata 30
<210> 437
<211> 23
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 437
gcaggaggaa aggaactatc tgg 23
<210> 438
<211> 30
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 438
ccatctcccc ctgctactat atattttatt 30
<210> 439
<211> 31
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 439
agagagacaa tgttttatga gacaaataag g 31
<210> 440
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 440
ttttcttagc attcaggtct cttttcaaa 29
<210> 441
<211> 29
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 441
ttctatgttc ttaaatgcag cttctacac 29
<210> 442
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 442
gcagcaaaag ccatcactgg 20
Claims (10)
1. A complex disease related SNP site primer composition is characterized in that: the SNP loci comprise: rs10229583, rs1048886, rs10811661, rs10814916, rs10886471, rs10906115, rs1111875, rs1116357, rs11257655, rs117601636, rs11787792, rs12000501, rs12010175, rs12549902, rs13266634, rs1359790, rs1421085, rs1436955, rs1470579, rs147538848, rs 0315505500, rs163171, rs 16316361329, rs 29955379, rs17584499, rs 177982, rs 18018095, rs1845900, rs187060802, rs1965305, rs 202773617640, rs 4120796, rs2206734, rs2237892, rs2237895, rs 223782237897, rs 22835637799, rs 70779, rs 70777943779, rs 7077563756375637563756375648, rs 70779, rs 7037563756375637563756375637563756375637569, rs 70729, rs 7037563756375637563756375637563756375637563756375637567, rs 703756375637563756375637563756375637563756375637567, rs 70729, rs 7037563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637567, rs1745, rs 70729, rs 707245, rs1745, rs 7245, rs 70729, rs 729, rs 70729, rs 707245, rs 7072563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756300, rs 729, rs1745, rs 703756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756300, rs1745, rs 729, rs1745, rs 7037563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756375637563756300, rs1745, rs 703756375637563756375637563756375637563756375637563756375637563756375637563756375637, rs10883795, rs11038167, rs11038172, rs1635, rs16887244, rs2142731, rs835784, rs10821944, rs 11900600673, rs12379034, rs 12617617617617656, rs13192471, rs1854853, rs2075876, rs2233434, rs2841277, rs2867461, rs3093024, rs3124, rs3781913, rs 65707075, rs10865331, rs13210693, rs17095830, rs30187, rs4552569, rs10036748, rs10845606, rs10847697, rs 2208334, rs1167796, rs1234315, rs12494314, rs 1287707, rs 13113177851338574, rs 1388190, rs1913517, rs 220793, rs 2205756375637563756375639, rs 31779, rs 70575648, rs 31575648 5648, rs 704746, rs 725646, rs 725648, rs 445648, rs 44575646, rs 445646, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 72563457435456345743545648, rs 725648, rs 445648, rs 725648, rs 445648, rs 725648, rs 43545648, rs 725648, rs 445648, rs 7256345743545648, rs 725648, rs 7256345743545648, rs 725648, rs 445634575648, rs 725648, rs 44563457435456345743545634575648, rs 445634575634575634575648, rs 445648, rs 7256345743545648, rs 445648, rs 435456345743545648, rs 445648, rs 725648, rs 445648, rs 725648, rs 43545648, rs 725648, rs 4456345743545648, rs 445648, rs 725648, rs 445648, rs 4354.
2. The primer composition of claim 1, wherein: the primer composition comprises:
3. the primer composition of claim 2, wherein: the complex disease comprises one or more of type 2 diabetes, hashimoto's thyroiditis, Alzheimer's disease, allergic dermatitis, rheumatoid arthritis, ankylosing spondylitis, systemic lupus erythematosus, asthma or schizophrenia.
4. Use of the primer composition of claim 1 in the preparation of a product for detecting and/or assessing a complex disease.
5. A product for detecting and/or assessing a complex disease, characterized in that: the product comprises the SNP site primer composition according to claim 1.
6. The product of claim 5, wherein: the product is an independent reagent or a kit.
7. Use of the primer composition of claim 1 or the product of claim 5 for detecting SNP sites associated with a complex disease.
8. A method for detecting SNP sites related to complex diseases, which is a non-disease diagnosis and treatment method and is characterized in that: the method comprises detecting SNP sites in the genome of a sample to be detected by using the primer composition of claim 1 or the product of claim 5.
9. The method of claim 8, wherein: the method comprises the following steps:
(1) extracting genome DNA;
(2) 1, performing multiplex PCR reaction;
(3) magnetic bead purification of round 1;
(4) performing multiplex PCR reaction of 2 nd round;
(5) magnetic bead purification for round 2;
(6) quantifying the library;
(7) detecting the quality of the library;
(8) performing computer sequencing on the library to obtain a fastaq sequence file, and performing quality evaluation, comparison and SNP (single nucleotide polymorphism) calling to obtain the genotype information of the SNP of the capture region of the sample to be detected;
(9) assigning the genotype of each SNP to the OR value of the disease in the population;
(10) multiplying all OR values of SNPs involved in a disease in an individual by the absolute genetic risk of the individual for the disease;
(11) the absolute genetic risk is ranked in a thousand human genome/Chinese human genome queue, namely the relative genetic risk of the individual aiming at the disease is ranked within 5% as low risk, above 95% as high risk and between 5 and 95% as normal range.
10. The method of claim 9, wherein: the quality evaluation software in the step (8) is FASTQC software, the comparison software is BWA software, and the SNP calling software is GATK software.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113846158A (en) * | 2021-11-24 | 2021-12-28 | 无锡中德美联生物技术有限公司 | Composite amplification kit for simultaneously detecting three chronic disease susceptibility genes and application thereof |
| CN114736963A (en) * | 2022-05-25 | 2022-07-12 | 首都医科大学宣武医院 | Substance combination for predicting Alzheimer disease onset risk, kit and application |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009146460A2 (en) * | 2008-05-30 | 2009-12-03 | Ordway Research Institute, Inc. | Methods for disease therapy |
| CN101845501A (en) * | 2010-05-18 | 2010-09-29 | 孟涛 | Comprehensive genetic analysis method of susceptibility of complex diseases |
| CN105112502A (en) * | 2015-07-07 | 2015-12-02 | 博奥颐和健康科学技术（北京）有限公司 | Complete primers for detecting SNP site combination associated with diabetes, and applications thereof |
| CN105200154A (en) * | 2015-11-02 | 2015-12-30 | 艾吉泰康生物科技(北京)有限公司 | Multiplex-PCR (polymerase chain reaction) detection method and kit for BRCA1 and BRCA2 gene mutation |
| CN105354444A (en) * | 2015-11-24 | 2016-02-24 | 华南理工大学 | Method for screening complex disease susceptible SNP combination based on susceptible SNP |
| CN109280695A (en) * | 2017-07-20 | 2019-01-29 | 浙江金华中科分数生命科技有限公司 | Utilize the polygenes score analysis method of human-body biological Samples Estimates complex disease onset risk |
| CN110499372A (en) * | 2019-09-18 | 2019-11-26 | 山西医科大学 | Multiplex PCR targeted capture parting system and kit based on high throughput sequencing technologies |
| CN112102884A (en) * | 2020-09-08 | 2020-12-18 | 苏州罗塞塔生物科技有限公司 | Multi-gene locus combined disease risk analysis and evaluation platform and method |
| CN112662766A (en) * | 2020-11-19 | 2021-04-16 | 赣南医学院第一附属医院 | Method for detecting familial thoracic aortic aneurysm and interlayer related mutant gene based on high-throughput sequencing technology |
-
2021
- 2021-06-11 CN CN202110651969.5A patent/CN113403380A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009146460A2 (en) * | 2008-05-30 | 2009-12-03 | Ordway Research Institute, Inc. | Methods for disease therapy |
| CN101845501A (en) * | 2010-05-18 | 2010-09-29 | 孟涛 | Comprehensive genetic analysis method of susceptibility of complex diseases |
| CN105112502A (en) * | 2015-07-07 | 2015-12-02 | 博奥颐和健康科学技术（北京）有限公司 | Complete primers for detecting SNP site combination associated with diabetes, and applications thereof |
| CN105200154A (en) * | 2015-11-02 | 2015-12-30 | 艾吉泰康生物科技(北京)有限公司 | Multiplex-PCR (polymerase chain reaction) detection method and kit for BRCA1 and BRCA2 gene mutation |
| CN105354444A (en) * | 2015-11-24 | 2016-02-24 | 华南理工大学 | Method for screening complex disease susceptible SNP combination based on susceptible SNP |
| CN109280695A (en) * | 2017-07-20 | 2019-01-29 | 浙江金华中科分数生命科技有限公司 | Utilize the polygenes score analysis method of human-body biological Samples Estimates complex disease onset risk |
| CN110499372A (en) * | 2019-09-18 | 2019-11-26 | 山西医科大学 | Multiplex PCR targeted capture parting system and kit based on high throughput sequencing technologies |
| CN112102884A (en) * | 2020-09-08 | 2020-12-18 | 苏州罗塞塔生物科技有限公司 | Multi-gene locus combined disease risk analysis and evaluation platform and method |
| CN112662766A (en) * | 2020-11-19 | 2021-04-16 | 赣南医学院第一附属医院 | Method for detecting familial thoracic aortic aneurysm and interlayer related mutant gene based on high-throughput sequencing technology |
Non-Patent Citations (9)
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113846158A (en) * | 2021-11-24 | 2021-12-28 | 无锡中德美联生物技术有限公司 | Composite amplification kit for simultaneously detecting three chronic disease susceptibility genes and application thereof |
| CN113846158B (en) * | 2021-11-24 | 2024-01-30 | 无锡中德美联生物技术有限公司 | Composite amplification kit for simultaneously detecting three chronic disease susceptibility genes and application thereof |
| CN114736963A (en) * | 2022-05-25 | 2022-07-12 | 首都医科大学宣武医院 | Substance combination for predicting Alzheimer disease onset risk, kit and application |
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