Ground mouse SNP (Single Nucleotide Polymorphism) molecular marker combination and application thereof

Abstract

The invention relates to the technical field of biological gene detection, and particularly discloses a hamster SNP (Single Nucleotide Polymorphism) molecular marker combination and application thereof. The hamster SNP molecular marker combination disclosed by the invention comprises at least one of 214 SNP molecular markers, and site information of the 214 SNP molecular markers is shown in a table 1. The hamster SNP molecular marker combination provided by the invention is used for detecting the SNP genotyping of the hamster, and has the properties of sensitivity, specificity, reliability, economy and high efficiency.

Description

Technical Field

[0001] The present application relates to the technical field of biological gene detection, and in particular to a hamster SNP molecular marker combination and application thereof. Background Art

[0002] The quality and physiological condition of experimental animals can directly affect the process of scientific research, product development, and the reliability of results in the production and testing of biological products. In life science research, the experimental animals used should have strict microbial control and clear and stable genetic background data for subsequent researchers to refer to. Therefore, reliable genetic quality monitoring plays an important role in ensuring animal quality and meeting requirements. The genetic quality monitoring of experimental animals uses scientific methods to conduct consistency tests on the genetic background of inbred animals and population genetics analysis of closed group animals to evaluate whether they meet the characteristics of the strain or group, and ensure the inherent genetic composition and biological characteristics of the breed and strain of animals. The genetic quality monitoring technology recommended in my country's national standards for experimental animals is relatively backward at present, and the detection rate, coverage, and judgment results are all inconsistent with the current needs of my country's comprehensive improvement of experimental animals. Commonly used genetic testing methods for experimental animals include morphological methods, immunological methods, biochemical markers, etc. With the rapid development of molecular biology, molecular genetic marker technology has been widely used in genetic testing of experimental animals. It is necessary to build an internationally advanced single nucleotide polymorphism (SNP) genetic quality monitoring method and standard technology system that can be used for conventional experimental animals and my country's special experimental animals in order to address the technical drawbacks of existing detection technology, such as backwardness, long time consumption, low accuracy and great harm to experimental animals.

[0003] Single nucleotide polymorphism (SNP) mainly refers to DNA sequence polymorphism caused by the variation of a single nucleotide (A, T, C and G) at the genomic level. SNP is widely present in biological genomes, with an average of 1 in every 1000 base pairs. SNP is a dimorphic marker caused by the conversion or transversion of a single base, which may be within the gene sequence or in the non-coding sequence outside the gene. Single nucleotide polymorphism (SNPS) is suitable for describing the genetic status of experimental animals such as mice due to its large number, wide distribution, distribution throughout the genome, and genetic stability. It has the advantages of non-invasive sampling, accuracy, time saving and suitability for automation. This technology aims to use single nucleotide polymorphism (SNP) markers to establish a cost-effective genetic quality detection system suitable for Chinese hamsters.

[0004] The mutation rate of SNP sites is about 10-8, which is only one ten-thousandth of the mutation rate of microsatellites. Compared with microsatellites, single nucleotide polymorphisms (SNPs) have the advantages of wide distribution, strong stability, and high throughput, and are more suitable as genetic monitoring markers. As a third-generation genetic marker, SNP detection technology has the advantages of accuracy, time saving, stable inheritance, and suitability for automation, and can be developed into SNP chips. Currently, SNP detection technology is being widely used in genetic evaluation of experimental animals, such as forensic identification, paternity testing, whole genome association analysis to find disease-related sites, animal genetic testing, and accelerated animal population directional breeding.

[0005] The Chinese hamster is a unique experimental animal resource in China, and it occupies an important position in life science research and is of great significance. Among rodents, the research on various biological traits and genomics of rats and mice is very in-depth and extensive, but there are few reports on basic data such as the genetic traits and genome of the Chinese hamster. At present, the Chinese hamster has been widely used in tumors, type II diabetes, lipid metabolism and parasites, and occupies an important position in life science research and is of great significance. However, there are relatively few methods for its genetic detection, and there are few reports on its basic data such as genetic traits and genome. At present, genetic detection is mainly carried out through microsatellite markers. Therefore, it is urgent and important to establish a SNP site genetic detection method for identifying Chinese hamster germplasm resources. Summary of the invention

[0006] In order to solve the above technical problems, the present application provides a hamster SNP molecular marker combination and application thereof.

[0007] In a first aspect, the present application provides a hamster SNP molecular marker combination, including at least one of 214 SNP molecular markers, and the site information of the 214 SNP molecular markers is shown in Table 1.

[0008] In view of the current lack of reliable data on the genetic genes of Chinese hamsters in my country, this application is to establish an economical and efficient genetic quality detection system for monitoring the genetic background of Chinese hamsters based on single nucleotide polymorphism (SNP) markers. By establishing scientific and rigorous screening and optimization steps, a SNP marker detection method suitable for Chinese hamsters was successfully established for the first time. The system has sensitivity, specificity, reliability and economy.

[0009] In a second aspect, the present application provides a method for screening the loci of the above-mentioned hamster SNP molecular marker combination, comprising the following steps: (1) Whole-genome sequencing and sequencing quality control of hamsters: The genomic DNA was genotyped using the DNBSEQ system combined with the probe-anchor synthesis method; the raw reads were filtered using fastp software; the paired ends were mapped to MunDraft using BWA software; SNP calling and filtering were performed using GATK4.1 software; the data were quality controlled using BCFtools software, and the following criteria were set: remove SNP sites with a depth of less than 2.0; remove SNP sites with a fisherman strand less than 60.0; remove SNP sites with an RMS mapping quality less than 30.0; and discard SNP sites with a depth less than 8.0; (2) Locus screening was performed according to the following five criteria: SNP loci were screened from different genomic fragments; the genotype frequency of the SNP loci was between 25% and 75%; there was no complete linkage between the SNP loci; no other SNPs or Indels were found within the 200 bp region upstream or downstream of the selected SNP loci; the SNP loci existed in the missense mutation, nonsense mutation and non-interrupted mutation regions of the gene region.

[0010] In a third aspect, the present application provides a primer set for detecting the above-mentioned hamster SNP molecular marker combination, and the position information of the primer set is shown in Table 4.

[0011] In a fourth aspect, the present application provides a hamster SNP detection kit, wherein the kit is used to detect the above-mentioned hamster SNP molecular marker combination.

[0012] Preferably, the kit comprises a primer set and / or a probe for detecting the hamster SNP molecular marker combination.

[0013] In a fifth aspect, the present application provides a hamster SNP gene detection system, including the hamster SNP molecular marker combination or the primer set of the hamster SNP molecular marker combination or the hamster SNP detection kit.

[0014] In a sixth aspect, the present application provides the application of at least one of the hamster SNP molecular marker combination, the primer set, the hamster SNP detection kit, and the hamster SNP gene detection system in the SNP genetic quality detection method and standard technical system of hamsters.

[0015] In the seventh aspect, the present application provides a method for detecting SNP markers in hamsters, comprising the following steps in sequence: collecting hamster samples; extracting and identifying the quality of hamster DNA; resequencing the whole genome of the hamster; comparing sequencing data, selecting sites with uniform genome distribution, good polymorphism, incomplete linkage, and high sequencing quality, to form a set of high-quality SNP site combinations; multiplex PCR primer design and quality verification of all SNP sites; forming a hamster multi-target region capture kit, and performing SNP site genotyping on all collected hamster individuals; multi-target region capture kit verification: sensitivity, repeatability, and specificity; first-generation sequencing to verify the typing accuracy of the kit; and population genetic structure analysis.

[0016] In summary, the technical solution of this application has the following effects: Based on single nucleotide polymorphism (SNP) markers, this application provides a combination of hamster SNP molecular markers and their corresponding applications, and establishes an economical and efficient genetic quality detection system for monitoring the genetic background of Chinese hamsters. By establishing scientific and rigorous screening and optimization steps, a SNP marker detection method suitable for Chinese hamsters was successfully established for the first time. The system has sensitivity, specificity, reliability and economy. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the specific experimental process of the hamster SNP gene detection system in this application.

[0018] Figure 2 This is the screening experimental process of SNP molecular markers in this application.

[0019] Figure 3 This is the principal component analysis diagram of 2 Chinese hamster populations with 55 SNPs in this application. DETAILED DESCRIPTION

[0020] The present application is further described in detail below in conjunction with embodiments. These embodiments should not be construed as limiting the scope of protection claimed in the present application. Example

[0021] This embodiment provides a hamster SNP gene detection system, the experimental process is as follows Figure 1 As shown, the specific steps are described as follows.

[0022] 1. Collection of Animal Samples All samples were from two populations of Shanxi Medical University: Chinese hamster population A: 31; Chinese hamster population B: 42. This experiment has been reviewed by the Animal Experimentation and Laboratory Animal Welfare Committee of Capital Medical University (AEEI-2021-309).

[0023] 2. Preparation of Hamster Genomic DNA The DNA was extracted from the ear tissue of Chinese hamster using Vazyme's FastPure Cell / Tissue DNA Isolation Mini Kit DC102. The absorbance A260 / A280 value was between 1.8 and 2.0 as detected by Nanodrop 2000c micro-spectrophotometer, and the DNA quality inspection was qualified, and it was stored at -20℃ for future use.

[0024] 3. Whole genome sequencing and sequencing quality control This application performed whole genome resequencing on 24 Chinese hamsters, which were derived from the Chinese hamster A population of Shanxi Medical University, and identified SNPs based on the Chinese hamster reference genome assembly (GCF_000223135.1).

[0025] The genomic DNA was genotyped using the DNBSEQ system combined with the probe-anchor synthesis method. The raw reads were filtered using fastp software. Paired-end mapping to MunDraft was performed using BWA software. SNP calling and filtering were performed using GATK4.1 software. The data were quality controlled using BCFtools software, and the following criteria were set: (1) SNP sites with a depth of less than 2.0 were removed; (2) SNP sites with a fisherman strand less than 60.0 were removed; (3) SNP sites with an RMS mapping quality less than 30.0 were removed; (4) SNP sites with a depth less than 8.0 were discarded.

[0026] 4.SNP locus screening The SNP locus screening criteria for this study include the following five points: 1. Screening of SNP loci from different genomic fragments; 2. The genotype frequency of the SNP locus is between 25% and 75%; 3. There is no complete linkage between SNP loci; 4. No other SNPs or Indels were found within the 200 bp region upstream or downstream of the selected SNP locus; 5. These SNP loci exist in the missense mutation, nonsense mutation and non-stop mutation regions of the gene region.

[0027] Through comparative analysis of the whole genome sequencing results of 24 Chinese hamsters, a total of 280,996 SNP sites were identified in this gene region, as the SNP sites in this gene region may be related to the genetic traits of the organism. According to the SNP screening standard process, this application screened out 214 high-quality SNP sites and constructed a Chinese hamster SNP detection system ( Figure 2 , Table 1).

[0028] Table 1 Information on 214 SNP loci in Chinese hamster 5. Kit Primer Design and Genotyping (1) After obtaining the candidate high-quality SNPs loci, the applicant will submit the region of interest (ROI), i.e., the physical location information of the SNPs, to Beijing Ajitaikang (Jiaxing) Biotechnology Co., Ltd.

[0029] (2) Using MFEprimer v3.1 software, highly specific multiplex PCR primers were designed at both ends of the ROI region based on thermodynamic stability; the length of the amplified sequence was in the range of 160 bp-260 bp.

[0030] (3) The quality of PCR primer pairs was verified, primer dimerization and nonspecific amplification were evaluated, the designed and synthesized primers were tested, and primers with poor detection effects were replaced.

[0031] (4) On the NovaSeqTM 6000 system, PE150 paired-end sequencing with multi-target region capture sequencing technology was used to perform genotyping of SNP loci.

[0032] (5) The instructions for the Hamster SNP Detection Kit are as follows: The first round of multiplex PCR reaction is: Take out the Enhancer buffer NB (1N), Primer pool, and IGT-EM808 polymerase mixture in the kit from the -20°C refrigerator in advance, place them on an ice box to melt, vortex briefly to mix, centrifuge them instantaneously, and place them on an ice box for later use.

[0033] Take out Enhancer buffer M from the 4°C refrigerator and thaw at room temperature.

[0034] Prepare the reaction system on an ice box according to Table 2: Table 2 First round multiplex PCR reaction system For the multiplex PCR reaction, the PCR instrument program was as follows: heating, 105°C; 95°C, 3 min 30 s; 98°C, 20 s; 55°C, 1 min; 60°C, 1 min; 65°C, 2 min; 72°C, 5 min; for 18 cycles.

[0035] Magnetic bead purification product: Prepare 80% ethanol with anhydrous ethanol and Nuclease-Free Water in advance and keep it at room temperature for later use. Please try to use freshly prepared 80% ethanol for magnetic bead purification.

[0036] Take out the purified magnetic beads from the 4°C refrigerator in advance, mix them and place them at room temperature for equilibration for 30 minutes; vortex the purified magnetic beads that have been equilibrated to room temperature and set aside.

[0037] Add 0.9 times the volume of magnetic beads (27 μL) to the 30 μL reaction system, pipette or vortex to mix, and let stand at room temperature for 5 minutes.

[0038] Centrifuge briefly and place the PCR tube on a magnetic rack for 3 minutes to wait for the solution to clarify.

[0039] Remove the supernatant completely, remove the PCR tube from the magnetic stand, add 50 μL YF buffer B to the tube, pipette to mix, and let stand at room temperature for 5 min.

[0040] Centrifuge briefly and place the PCR tube on the DynaMag-96 Side magnetic stand for 3 minutes.

[0041] Keep the PCR tube on the magnetic rack, carefully remove the supernatant, add 180 μL 80% ethanol solution into the PCR tube, and let it stand for 30 seconds.

[0042] Keep the PCR tube on the magnetic rack, discard the supernatant, add 180 μL 80% ethanol solution to the PCR tube again, let it stand for 30 seconds, and discard the supernatant.

[0043] Put the cap on the tube and centrifuge briefly to remove the residual ethanol to the bottom of the tube. Place the PCR tube on the magnetic rack and carefully use a 10μL pipette to remove the residual ethanol at the bottom, being careful not to suck up the magnetic beads.

[0044] Keep the PCR tube on the magnetic rack and let it stand at room temperature for 3 to 5 minutes to dry the magnetic beads and allow the residual ethanol to evaporate completely.

[0045] Add 24 μL Nuclease-Free Water, remove the PCR tube from the magnetic stand, pipette or vortex to mix, and let stand at room temperature for 2 minutes.

[0046] Centrifuge briefly and place the PCR tube on a magnetic rack for 2 minutes to wait for the solution to clarify.

[0047] Use a pipette to take up 13.5 μL of supernatant and transfer it to a new PCR tube. The supernatant in the tube is the purified multiple PCR product, which is marked and used for PCR reaction.

[0048] The second round of adapter sequence PCR reaction is: Reaction system: Prepare the reaction system according to Table 3 on an ice box: Table 3 Second round of linker sequence PCR reaction system Reactants Volume (μL) Multiplex PCR products purified in the previous step 13.5 Enhancer buffer M 2.5 Premixed adapter primers, concentration 10 μM each 1 IGT-EM808polymerase mixture 10 ddH2O 3 Total 30 The reaction conditions were as follows: heating at 105°C, 95°C for 3 min 30 s, 98°C for 20 s, 58°C for 1 min, 72°C for 30 s, and 72°C for 5 min for 9 cycles.

[0049] Second round of magnetic bead purification: Prepare 80% ethanol with anhydrous ethanol and Nuclease-Free Water in advance and keep it at room temperature for later use. Please try to use freshly prepared 80% ethanol for magnetic bead purification.

[0050] Take out the purified magnetic beads from the 4°C refrigerator in advance, mix them and place them at room temperature for equilibration for 30 minutes; vortex the purified magnetic beads that have been equilibrated to room temperature and set aside.

[0051] Add 0.9 times the volume of magnetic beads (27 μL) to the 30 μL reaction system, pipette or vortex to mix, and let stand at room temperature for 5 minutes.

[0052] Centrifuge briefly and place the PCR tube on a magnetic rack for 3 minutes to wait for the solution to clarify.

[0053] Remove the supernatant completely, remove the PCR tube from the magnetic stand, add 50 μL YF buffer B to the tube, pipette to mix, and let stand at room temperature for 5 min.

[0054] Centrifuge briefly and place the PCR tube on the DynaMag-96 Side magnetic stand for 3 minutes.

[0055] Keep the PCR tube on the magnetic rack, carefully remove the supernatant, add 180 μL 80% ethanol solution into the PCR tube, and let it stand for 30 seconds.

[0056] Keep the PCR tube on the magnetic rack, discard the supernatant, add 180 μL 80% ethanol solution to the PCR tube again, let it stand for 30 seconds, and discard the supernatant.

[0057] Put the cap on the tube and centrifuge briefly to remove the residual ethanol to the bottom of the tube. Place the PCR tube on the magnetic rack and carefully use a 10μL pipette to remove the residual ethanol at the bottom, being careful not to suck up the magnetic beads.

[0058] Keep the PCR tube on the magnetic rack and let it stand at room temperature for 3 to 5 minutes to dry the magnetic beads and allow the residual ethanol to evaporate completely.

[0059] Add 24 μL Nuclease-Free Water, remove the PCR tube from the magnetic stand, pipette or vortex to mix, and let stand at room temperature for 2 minutes.

[0060] Centrifuge briefly and place the PCR tube on a magnetic rack for 2 minutes to wait for the solution to clarify.

[0061] Use a pipette to take 20 μL of supernatant and transfer it to a new PCR tube. The supernatant in the tube is the prepared multiplex PCR library.

[0062] Take 1 μL of library for concentration measurement and record the library concentration (the concentration range of normal library is 20 ng / μL to 40 ng / μL, and the library concentration is mainly related to the quality of the template.

[0063] (6) The quality of Chinese hamster PCR primers was verified, primer dimerization and nonspecific amplification were evaluated, the designed and synthesized primers were tested, and primers with poor detection effects were replaced. Ultimately, the matching degree of 214 primers on the genome reached 100%. The length of the amplified fragment ranged from 160 bp to 260 bp (Table 4).

[0064] Table 4 Positions and predicted coverage of PCR primers in Chinese hamster 6. Validation of Multi-target Region Capture Sequencing Kit The Chinese hamster SNP site genetic detection kit was simultaneously verified by three companies (Aijitaikang, Tianyi Huiyuan, and Zhongke Guobang). As shown in Table 5 below, the sensitivity, specificity, and repeatability of the Chinese hamster kit were verified: Table 5 Validation of Chinese Hamster Kit The experimental results of the three units showed that when the concentration of Chinese hamster DNA was as low as 10 μg / mL, the kit could still perform multiple PCR amplification of the SNP combination of Chinese hamsters and accurately type the SNP loci, which showed that the sensitivity of the Chinese hamster kit was good; the typing results of the SNP loci of the four Chinese hamster DNA samples with a concentration of 40 μg / mL were completely consistent, which showed that the Chinese hamster kit had strong repeatability (Table 6); the experiment used 40 μg / mL miniature pig DNA to verify the specificity of the Chinese hamster kit and found that the Chinese hamster kit could not amplify and type the SNP loci of miniature pig DNA samples, which showed that the specificity of the Chinese hamster kit was good. In summary, the Chinese hamster kit performed well in terms of sensitivity, specificity and repeatability.

[0065] Table 6 Validation results of Chinese hamster test kit 7. Population genetic structure analysis By performing SNP typing tests on the collected samples, the present application optimized the sites. The MAPQ (mapping quality) values ​​of 4 SNP sites (CH48, CH65, CH100 and CH139) were less than 59, indicating that the amplified fragments of these 4 SNP sites did not uniquely match the reference genome, and the genotyping results were unreliable. The present application also found that 73 SNP sites (such as CH12, CH16, CH17, etc.) were monomorphic in all samples, so the present application believes that these SNP sites with poor polymorphism are not suitable for population analysis. In addition, there are 82 SNP sites (such as CH43, CH188, CH189, etc.) in the SNP panel that are completely linked, and it is believed that these SNP sites have the same therapeutic effect, so one SNP site is retained for each group of completely linked SNP sites. Finally, 55 high-quality SNP sites were optimized, and these 55 SNP sites were recommended for genetic evaluation of the Chinese hamster population.

[0066] The application uses popgene32 software to analyze the genetic structure (table 7) of two Chinese hamster populations. In the remaining 55 SNP sites, CH183 and CH195 show monomorphism in A population. CH51, CH51, CH89 and CH105 show monomorphism in B population. By using 55 SNP sites to analyze two populations of Chinese hamster, it is found that the average number of alleles observed, the average number of effective alleles, the average Shannon index I, the average heterozygosity Obs Het observed, the average heterozygosity Exp Het expected to be higher than the average heterozygosity Ave Het of Chinese hamster A population are all higher than the Chinese hamster B population. Supplementary table shows the relevant parameters of two populations of Chinese hamster.

[0067] Among the 55 SNP sites in the Chinese hamster population A, 53 sites showed dialleles. The average observed allele number of population A was 1.9636, the average effective allele number was 1.7146, the average heterozygosity was 0.3945, the average polymorphism information content was 0.3076, there were 0 highly polymorphic sites (PIC>0.5), 46 moderately polymorphic sites (0.25<PIC<0.5), and 7 lowly polymorphic sites (PIC<0.25). Among the 55 SNP loci, 51 loci showed bi-allelic genotypes in the Chinese hamster population B. The average number of observed alleles in population B was 1.9273, the average number of effective alleles was 1.6252, the average heterozygosity was 0.3754, the average polymorphism information content was 0.3492, there were 0 highly polymorphic loci (PIC>0.5), 42 moderately polymorphic loci (0.25<PIC<0.5), and 9 lowly polymorphic loci (PIC<0.25). Among the 55 SNP loci, there were 37 loci with higher average polymorphism information content in population A than in population B.

[0068] Table 7 Genetic structure of two Chinese hamster populations This application also conducted a differential SNP analysis of two Chinese hamster populations. Through the chi-square test, CH1, CH62, The seven SNP loci CH71, CH92, CH96, CH117, and CH122 are differential SNP loci between population A and population B. In these SNP loci, the genotype frequencies of the two populations are different. Table 8 Differential loci of Chinese hamsters

[0069] Table 8 Differential SNP loci between two populations of Chinese hamsters 8. PCA Principal Component Analysis PCA analysis was performed on the Chinese hamster population using 55 SNP loci and differential SNP loci. By reducing the dimension of the data, PCA can be calculated to show the main differences in genetic distance between complex samples. The PCA results based on the allele frequency distribution of 55 SNP loci and 7 differential loci in the Chinese hamster population are shown in the figure. Figure 2 In the PCA results based on genotype, the two populations of Chinese hamsters are difficult to separate ( Figure 3 , the left figure is the principal component analysis of 55 SNP sites, and the right figure is the PCA result of the allele frequency distribution of 7 differential sites). This indicates that the two populations of Chinese hamsters may have come from the same population.

[0070] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A hamster SNP molecular marker combination, characterized in that: Including at least one of 214 SNP molecular markers, the site information of the 214 SNP molecular markers is shown in Table 1: Table 1 Information on 214 SNP loci in Chinese hamster 2. The method for screening loci of a hamster SNP molecular marker combination according to claim 1, characterized in that: The following steps are involved: (1) Whole-genome sequencing and sequencing quality control of hamsters: The genomic DNA was genotyped using the DNBSEQ system combined with the probe-anchor synthesis method; the raw reads were filtered using the fastp software; the paired ends were mapped to MunDraft using the BWA software; SNPs were called and filtered using the GATK4.1 software; the data were quality controlled using the BCFtools software, and the following criteria were set: SNP sites with a depth of less than 2.0 were removed; SNP sites with a fisherman strand less than 60.0 were removed; SNP sites with an RMS mapping quality less than 30.0 were removed; SNP sites with a depth less than 8.0 were discarded; (2) Locus screening was performed according to the following five criteria: SNP loci were screened from different genomic fragments; the genotype frequency of the SNP loci was between 25% and 75%; there was no complete linkage between the SNP loci; no other SNPs or Indels were found within the 200 bp region upstream or downstream of the selected SNP loci; the SNP loci existed in the missense mutation, nonsense mutation and non-interrupted mutation regions of the gene region.

3. A primer set for detecting the hamster SNP molecular marker combination according to claim 1, characterized in that: The position information of the primer set is shown in Table 4: Table 4 Positions and predicted coverage of PCR primers in Chinese hamster 4. A hamster SNP detection kit, characterized in that: The kit is used to detect the hamster SNP molecular marker combination according to claim 1.

5. The hamster SNP detection kit according to claim 4, characterized in that: The kit comprises a primer set and / or a probe for detecting the hamster SNP molecular marker combination according to claim 1.

6. A hamster SNP gene detection system, characterized in that: The invention comprises the hamster SNP molecular marker combination according to claim 1 or the primer set of the hamster SNP molecular marker combination according to claim 3 or the hamster SNP detection kit according to any one of claims 4-5.

7. The hamster SNP gene detection system according to claim 6, characterized in that: The method comprises the following steps in sequence: collecting hamster samples; extracting and authenticating hamster DNA; resequencing the whole genome of hamsters; comparing sequencing data, selecting sites with uniform genome distribution, good polymorphism, incomplete linkage and high sequencing quality to form a set of high-quality SNP site combinations; designing and quality verifying multiplex PCR primers for all SNP sites; forming a hamster multi-target region capture kit to perform SNP site genotyping on all collected hamster individuals; validating the sensitivity, repeatability and specificity of the multi-target region capture kit; validating the typing accuracy of the kit through first-generation sequencing; and analyzing the genetic structure of the population.

8. Use of at least one of the hamster SNP molecular marker combination according to claim 1, the primer set according to claim 3, the hamster SNP detection kit according to claim 4, and the hamster SNP gene detection system according to any one of claims 6 to 7 in a hamster SNP genetic quality detection method and standard technical system.

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