SNP (Single Nucleotide Polymorphism) site combination and method for identifying or assisting in identifying mouse genetic background

Abstract

The invention provides an SNP (Single Nucleotide Polymorphism) site combination and a method for identifying or assisting in identifying a mouse genetic background. The invention provides an SNP (Single Nucleotide Polymorphism) site combination for identifying or assisting in identifying the genetic background of a mouse. The SNP site combination totally comprises 32 SNP sites. The invention provides a method for identifying or assisting in identifying the genetic background of the mouse on the basis of the SNP site combination, and the genotype of the SNP site combination of the mouse to be identified is matched with the reference gene spectrum of the SNP site combination of the target strain of mouse, so that the identification or auxiliary identification of the genetic background of the mouse to be identified is realized. The method provided by the invention has the characteristics of high cost performance, high identification speed and high accuracy, and can meet the requirements of low-cost and high-accuracy mouse genetic quality control.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a SNP site combination and method for identifying or assisting in identifying the genetic background of mice. Background Art

[0002] Genetic quality monitoring is an important part of the quality control of experimental animals, and a clear genetic background is a prerequisite to ensure that scientific research is reliable and accurate. Traditional genetic background detection methods for experimental animals include phenotypic detection methods such as immune markers and biochemical markers, but they have limitations and cannot accurately reflect the genetic background of experimental animals. With the development of science and technology, DNA detection technology has begun to be widely used. SNP, as a third-generation genetic marker, has the advantages of large number and wide distribution, high genotype, strong representativeness and easy analysis. Providing a SNP site combination for identifying the genetic background of mice is a technical problem that needs to be solved urgently in this field. Summary of the invention

[0003] The present invention provides a SNP site combination for identifying or assisting in identifying the genetic background of mice.

[0004] In a first aspect, the present invention provides an application of a material for detecting the genotype of a SNP site combination in identifying or assisting in identifying the genetic background of a mouse, wherein the SNP site combination includes 32 SNP sites shown in Table 1:

[0005] Table 1 Combination of 32 SNP loci

[0006]

[0007]

[0008] In Table 1, the rs3022953, rs3023442, and rs3023450 loci were based on the GRCm38 version, and the remaining loci were based on the GRCm39 version.

[0009] As described above, the material for detecting the genotype of the SNP site combination includes an amplification primer composition and a single-base extension composition for amplifying 32 SNP sites; the amplification primer composition includes a single-stranded DNA molecule with a nucleotide sequence of SEQ ID NO: 1-64; the single-base extension composition includes a single-stranded DNA molecule with a nucleotide sequence of SEQ ID NO: 65-96.

[0010] In a second aspect, the present invention provides an amplification primer composition for detecting the genotype of the above-mentioned SNP site combination, comprising a single-stranded DNA molecule with a nucleotide sequence of SEQ ID NO: 1-64.

[0011] In a third aspect, the present invention provides a single-base extension composition for detecting the genotype of the above-mentioned SNP site combination, comprising a single-stranded DNA molecule having a nucleotide sequence of SEQ ID NO: 65-96.

[0012] In a fourth aspect, the present invention provides a product for detecting the genotype of the above-mentioned SNP site combination, comprising the above-mentioned amplification primer composition and / or single-base extension composition.

[0013] The product as described above may be a kit.

[0014] The product as described above may also include other reagents required for detecting the genotype of the above SNP site combination, for example, it may include DNA polymerase and dNTP required for PCR, and may also include SAP enzyme (Shrimp Alkaline Phosphatase) and the like.

[0015] In a fifth aspect, the present invention provides the use of the above-mentioned amplification primer composition or the above-mentioned single-base extension composition or the above-mentioned product in identifying or assisting in identifying the genetic background of mice.

[0016] In a sixth aspect, the present invention provides a method for identifying or assisting in identifying the genetic background of a mouse, comprising:

[0017] Obtaining genomic DNA of the mouse to be identified;

[0018] Detecting the genotype of the above SNP site combination in the mouse genomic DNA sample to be identified;

[0019] The genotype of the SNP site combination of the target strain of mice is used as the standard gene spectrum, and the genotype of the SNP site combination of the mouse to be identified is compared with the standard gene spectrum to identify or assist in identifying the genetic background of the mouse to be identified.

[0020] In the above method, the mouse strain refers to a group of mice with similar genetic characteristics under a specific genetic background. Different mouse strains have different genetic backgrounds. The target strain refers to mice with a known genetic background and a determined strain.

[0021] In the method described above, the mouse is one of an inbred strain, a congenic strain or a hybrid group, which can be specifically determined according to experimental needs; in the present invention, the inbred strain mouse can be, for example, at least one of 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB, and SJL; the congenic strain mouse can be, for example, at least one of SCID-Beige, B6 Albino, APOE, and CB-17SCID; the hybrid group mouse can be, for example, at least one of CB6F1 and B6D2F1.

[0022] As described above, the mouse strain is defined according to the national standard of the People's Republic of China numbered GB-14923-2022. Specifically, an inbred mouse refers to a strain in which more than 98.6% of the loci in the genome of any individual in the mouse population are pure; a congenic strain refers to an inbred strain formed by backcrossing by selecting a specific marker from a donor strain, which differs from the original inbred strain only in a very small chromosome fragment; a hybrid group refers to a group of offspring produced by hybridization of two different inbred strains.

[0023] In the method described above, the mouse to be identified can be one of an inbred strain, a congenic strain or a hybrid group. When the mouse to be identified is inferred to be a certain inbred strain or a hybrid group, the corresponding target strain mouse selects the same inbred strain or hybrid group, and when the genotypes of the above SNP sites of the two are consistent, it means that the genetic background of the mouse to be identified is the same as that of the target strain mouse; when the mouse to be identified is inferred to be a congenic strain, if the genotype of the above SNP sites of the mouse to be identified is the same as that of the target strain mouse, it is inferred that the mouse to be identified is derived from the genetic background of the target strain mouse.

[0024] In the method described above, the genotype of the SNP site combination of the target strain of mice can be obtained from a database, or by testing the genotype of the SNP site combination of known strains of mice; wherein the database can be the National Center for Biotechnology Information (NCBI) or Mouse Phenome Datebase.

[0025] The method as described above, detecting the genotype of the SNP site combination in the mouse genomic DNA sample to be identified, specifically comprises:

[0026] Using the DNA of the mouse to be identified as a template, multiplex PCR amplification is performed using an amplification primer combination for amplifying the 32 SNP sites to obtain an amplification product;

[0027] Using SAP enzyme to perform enzymatic digestion reaction on the amplified product to obtain an enzymatic digestion product;

[0028] Using the single-base extension composition of the 32 SNP sites to extend the enzymatic digestion product to obtain an extension product;

[0029] The extension products were analyzed by mass spectrometry using MALDI-TOF, and the genotype of the SNP site combination was obtained based on the analysis results.

[0030] In a seventh aspect, the present invention provides a method for detecting or assisting in detecting the genetic quality of mice, comprising:

[0031] Obtain genomic DNA of the mouse to be tested;

[0032] Detecting the genotype of the above SNP site combination in the mouse genomic DNA sample to be identified;

[0033] The genotype of the SNP site combination of the mouse to be identified is compared with the standard genotype of the SNP site combination of the target strain mouse. When the genotype of the SNP site combination of the mouse to be identified is the same as the standard genotype of the SNP site combination of the target strain mouse, the genetic quality is judged to be stable, otherwise the genetic quality is unqualified.

[0034] The present invention provides a SNP site combination for identifying or assisting in identifying the genetic background of mice, and provides a method for identifying or assisting in identifying the genetic background of mice based on the SNP site combination. According to the method provided by the present invention, the genetic background of the mouse to be identified is finally confirmed by matching the genotype of the SNP site combination of the mouse to be identified with the genotype of the SNP site combination of the target strain mouse. The method provided by the present invention has the characteristics of high cost performance, fast identification speed and high accuracy, so as to meet the requirements of low-cost and high-accuracy monitoring of mouse genetic quality control. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 This is the MALDI-TOF mass spectrum of the rs3022883 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0036] Figure 2 This is the MALDI-TOF mass spectrum of the rs3023034 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0037] Figure 3 This is the MALDI-TOF mass spectrum of the rs3023039 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0038] Figure 4This is the MALDI-TOF mass spectrum of the rs3023064 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0039] Figure 5 This is the MALDI-TOF mass spectrum of the rs3023203 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0040] Figure 6 This is the MALDI-TOF mass spectrum of the rs3023226 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0041] Figure 7 This is the MALDI-TOF mass spectrum of the rs3023251 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0042] Figure 8 It is the MALDI-TOF mass spectrum of the rs3023342 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0043] Fig. 9 This is the MALDI-TOF mass spectrum of the rs3023381 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0044] Fig.10 This is the MALDI-TOF mass spectrum of the rs3023382 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0045] Fig.11 This is the MALDI-TOF mass spectrum of the rs3023442 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0046] Fig.12 This is the MALDI-TOF mass spectrum of the rs3023481 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0047] Fig.13 This is the MALDI-TOF mass spectrum of the rs3088673 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0048] Fig.14 This is the MALDI-TOF mass spectrum of the rs3089604 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0049] Fig.15This is the MALDI-TOF mass spectrum of the rs3091174 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0050] Fig.16 This is the MALDI-TOF mass spectrum of the rs3659787 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0051] Fig.17 This is the MALDI-TOF mass spectrum of the rs3702158 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0052] Fig.18 This is the MALDI-TOF mass spectrum of the rs3706082 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0053] Fig.19 This is the MALDI-TOF mass spectrum of the rs3709624 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0054] Fig. 20 This is the MALDI-TOF mass spectrum of the rs3722313 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0055] Fig.21 This is the MALDI-TOF mass spectrum of the rs13481123 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0056] Fig. 22 This is the MALDI-TOF mass spectrum of the rs13481154 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0057] Fig.23 This is the MALDI-TOF mass spectrum of the rs3022796 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0058] Fig.24 This is the MALDI-TOF mass spectrum of the rs3022825 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0059] Fig.25 This is the MALDI-TOF mass spectrum of the rs3022953 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0060] Fig.26This is the MALDI-TOF mass spectrum of the rs3022977 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0061] Fig. 27 This is the MALDI-TOF mass spectrum of the rs3023177 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0062] Fig.28 This is the MALDI-TOF mass spectrum of the rs3023436 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0063] Fig.29 It is the MALDI-TOF mass spectrum of the rs3023450 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0064] Fig.30 This is the MALDI-TOF mass spectrum of the rs3089349 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0065] Fig.31 This is the MALDI-TOF mass spectrum of the rs3089984 site of the mouse numbered 20240910-6 (C57BL / 6N);

[0066] Fig.32 This is the MALDI-TOF mass spectrum of the rs3724876 site of the mouse numbered 20240910-6 (C57BL / 6N).

[0067] Figure 1-Figure 32 In the figure, the peak corresponding to the blue line is the judgment peak. DETAILED DESCRIPTION

[0068] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings in the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments, and they should not be understood as limitations on the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. In the description of the present invention, it should be understood that the terms used are only for descriptive purposes and cannot be understood as indicating or implying relative importance.

[0069] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0070] Example 1

[0071] 1. 32 SNP loci combination and substances for detecting 32 SNP loci combination

[0072] The 32 SNP site combinations provided by the present invention and the amplification primers and single-base extension primers required for detecting the SNP sites are shown in Table 2.

[0073] Table 2 Combinations of 32 SNP loci and their detection primers

[0074]

[0075]

[0076]

[0077] 2. Establishment of SNP site combination genotype detection method

[0078] 2.1. Synthesis of amplification primers and extension primers required for 32 SNP sites

[0079] The amplification primers shown in Table 2 were synthesized, and the synthesized single amplification primers were redissolved in deionized water to obtain a storage solution with a concentration of 100 μmol / L. The storage solutions of 64 amplification primers corresponding to the 32 SNP site combinations were mixed in equal volumes, and deionized water was added to make up the volume to obtain an amplification primer composition in which the final concentration of each amplification primer was 0.5 μmol / L, which was named amplification primer Mix.

[0080] The extension primers shown in Table 2 were synthesized. According to the relative molecular mass and OD of the primers, the target reaction concentration (μM) of each primer was calculated as LN (Extension Primer Mass)-7.82, the target working concentration (μM) = (target reaction concentration * 9) / 0.94) * the number of extension primers mixed (such as 15 extension primers mixed, the number is 15), and the amount of deionized water to be added to each primer tube (μL) = extension primer OD * 33 / extension primer Mass / target working concentration * 1000000 was calculated. The primers were redissolved according to the calculated amount to obtain the extension primer storage solution. An equal volume of the extension primer storage solution was taken and fully mixed to obtain a single-base extension composition, which was named Extension Primer Mix.

[0081] 2.2. Testing of genotypes of SNP loci combinations of 16 common mouse strains

[0082] 2.2.1. From breeding populations with clear genetic backgrounds, mice of 10 inbred strains, including 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB and SJL, mice of 2 hybrid strains, including CB6F1 and B6D2F1, mice of 4 congenic strains, including SCID-Beige, B6 Albino, APOE and CB-17SCID, were collected. Three mice were randomly selected from each strain. The tails of 48 mice were collected, and the length of the tails was not less than 0.5 cm.

[0083] 2.2.2. Use the fully automatic nucleic acid extraction instrument and ABI magnetic bead nucleic acid extraction kit to extract genomic DNA from mouse tails to obtain a template DNA solution with a concentration of 10-20 ng / μL. Pipette 20 μL of the template DNA solution into a 96-well plate and arrange them in sequence, marking them for later use.

[0084] 2.2.3. Use the PCR reagents provided by Agena Company, prepare the reagent amount 10% more than the number of tests according to the PCR reaction system shown in Table 3, add the prepared mixture to the PCR plate wells at a volume of 4 μL / well, and finally add 1 μL / well of template DNA to obtain a PCR reaction system with a total volume of 5 μL / well. Perform PCR amplification according to the PCR reaction program shown in Table 4 to obtain the amplified product.

[0085] Table 3 PCR reaction system

[0086] Components Final concentration Volume (μL) water / 1.8 Amplification primer Mix (0.5 μM) 0.1μM 1 <![CDATA[10*Buffer (containing Mg 2+ )]]> <![CDATA[2mM MgCl2]]> 0.5 <![CDATA[MgCl2(25mM)]]> 2mM 0.4 dNTP(25mM) 500μM 0.1 PCR enzymes 1Unit 0.2 Template DNA (10 ng / μL) 1 Total 5

[0087] Table 4 PCR reaction procedure

[0088]

[0089] 2.2.4 Enzymatic hydrolysis

[0090] Use the Agena iPLEX Gold kit and the SAP reaction system shown in Table 5 to prepare a mixture 10% more than the detection amount, named SAP enzyme Mix, add 2 μL / well of the prepared SAP enzyme Mix to each amplification product obtained in 2.2.3, pipette and mix well, perform SAP enzymatic reaction in a PCR instrument according to the enzymatic reaction program shown in Table 6 to obtain enzymatic products.

[0091] Table 5 SAP reaction system

[0092] Components Volume (μL) SAP Buffer 0.17 SAP enzyme 0.3 water 1.53 Total 2

[0093] Table 6 SAP enzymatic hydrolysis reaction procedure

[0094]

[0095]

[0096] 2.2.5. Single base extension reaction

[0097] Using the iPLEX Gold kit from Agena, a mixture was prepared according to the reaction system shown in Table 7 in an amount 10% more than the number of tests, named single base extension reaction Mix, 2 μL / well of the prepared single base extension reaction Mix was added to the above enzymatic hydrolysis product, pipetted and mixed, and a single extension reaction was performed in a PCR instrument according to the reaction program shown in Table 8 to obtain an extension product.

[0098] Table 7 Single base extension reaction system

[0099] Components Volume (μL) UEP Primer Mix 0.94 Gold Buffer 0.2 Stop Buffer 0.1 iPlex Enzymes 0.04 water 0.62 Total 2

[0100] Table 8 Single base extension reaction program

[0101]

[0102] 2.2.6 Mass spectrometry

[0103] The extension product was desalted and purified, and the purified product was transferred to the corresponding matrix point of the chip using the MassARRAY Nanodispenser RS1000 device, and detected using the MassARRAY Analyzer to obtain the detection genotypes of 48 mouse samples at 32 SNP sites. The test results are shown in Table 9. The MALDI-TOF mass spectra of the 32 SNP sites of the mouse with the sample ID of 20240910-6 (C57BL / 6N) are shown in Figure 1-32 shown.

[0104] There were 9 sites with no call in the 48 samples, including rs3023064 of sample 20240910-48, rs3089604 and rs3722313 of sample 20240910-22, rs3089604, rs3722313 and rs3023064 of sample 20240910-38, rs3022796, rs3022977 and rs13481123 of sample 20240910-21. The genotypes shown in Table 9 were manually interpreted based on the mass spectrometry peaks. (When the no call results were concentrated in a certain sample and other samples were normal, it was considered to be caused by the quality of the nucleic acid of the sample.) The lowest call rate of the sample was 90.6% and the highest was 100%. All met more than 90%, and the detection rate was good.

[0105] According to Table 9, the genotypes of the 32 SNP sites detected by the method provided by the present invention as shown in Table 9 are compared with the reference gene spectrum of mice. It can be seen that the detection results shown in Table 9 match the actual situation 100%, indicating that the accuracy of the method provided by the present invention is high. The genotype detection results of the 32 SNP sites of the 16 strains of mice provided by the present invention can be distinguished from each other, indicating good specificity. The detection genotypes of the 32SNPs of 3 different samples of the same strain provided by the present invention are consistent (except BALB / c Nude), indicating good stability.

[0106] 3. Comparison of SNP genotypes of 16 mouse strains with reference gene maps

[0107] In the Mouse Phenome Database, search for 129S1 / Sv ImJ, 129S2 / Sv Hsd, BALB / cByJ, BALB / cJ, C3H / He H, C3H / He J, C3H / He SnJ, C57BL / 6J, C57BL / 6J Crl, C57BL / 6N Crl, C57BL / 6NJ, DBA / 1J, DBA / 2J, FVB / NJ, SJL / J, NOD.Cg-Prkdc scid II tm1WjI The genotypes of 32 SNP loci of / SzJ (NOG), NOD / Shi Lt J (NOD SCID), and BALB / c nude strain mice were used as reference gene maps as shown in Table 10. The reference gene maps of hybrid mice CB6F1 and B6D2F1 were obtained by combining the standard genotypes of the parent strains.

[0108] The combined genotypes of the SNP sites of the 16 strains shown in Table 9 were compared and calculated with the reference gene map shown in Table 10. If the SNP sites of the 16 strains of mice were consistent with the genotypes of the reference gene map, the gene match number was recorded as 1, otherwise it was recorded as 0. The numerical value corresponding to the 32 SNP sites of each sample was calculated = gene match number / 32*100%. The calculation results are shown in Table 11. According to Table 11, the test results of inbred and hybrid mice match the standard gene spectrum of the corresponding strains of mice 100%, indicating that the method provided by the present invention can distinguish and identify these inbred and hybrid mice; the sample test results of SCID-Beige and CB-17SCID strains match the BALB / cJ reference gene spectrum 100%, the sample test results of B6 Albino match the C57BL / 6NJ reference gene spectrum 100%, and the sample test results of APOE match the C57BL / 6J reference gene spectrum 100%, indicating that congenic strains cannot be successfully distinguished, but the background sources of these congenic strains can be clarified.

[0109] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

Claims

1. Use of a substance for detecting the genotype of a combination of SNP sites in identifying or assisting in identifying the genetic background of mice, characterized in that: The SNP site combination includes 32 SNP sites shown in Table 1: Table 1 Combination of 32 SNP loci In Table 1, the rs3022953, rs3023442, and rs3023450 loci were based on the GRCm38 version, and the remaining loci were based on the GRCm39 version.

2. The use according to claim 1, characterized in that: The material for detecting the genotype of the SNP site combination includes an amplification primer composition and a single-base extension composition for amplifying 32 SNP sites; the amplification primer composition includes a single-stranded DNA molecule with a nucleotide sequence of SEQ ID NO: 1-64; the single-base extension composition includes a single-stranded DNA molecule with a nucleotide sequence of SEQ ID NO: 65-96.

3. An amplification primer composition for detecting the genotype of the SNP site combination according to claim 1, characterized in that: It includes a single-stranded DNA molecule having a nucleotide sequence of SEQ ID NO: 1-64.

4. A single base extension composition for detecting the genotype of the SNP site combination according to claim 1, characterized in that: The single-stranded DNA molecule comprises the nucleotide sequence of SEQ ID NO: 65-96.

5. A product for detecting the genotype of the SNP site combination according to claim 1, characterized in that: It comprises the amplification primer composition according to claim 3 and / or the single base extension composition according to claim 4.

6. Use of the amplification primer composition according to claim 3, the single-base extension composition according to claim 4, or the product according to claim 5 in identifying or assisting in identifying the genetic background of mice.

7. A method for identifying or assisting in identifying the genetic background of mice, characterized in that: include: Obtain genomic DNA of the mouse to be identified; Detecting the genotype of the SNP site combination according to claim 1 in the mouse genomic DNA sample to be identified; The genotype of the SNP site combination of the target strain of mice is used as the standard gene spectrum, and the genotype of the SNP site combination of the mouse to be identified is compared with the standard gene spectrum to identify or assist in identifying the genetic background of the mouse to be identified.

8. The method according to claim 7, characterized in that The mouse is one of an inbred strain, a congenic strain or a hybrid group.

9. The method according to claim 7, characterized in that: Detecting the genotype of the SNP site combination of claim 1 in the mouse genomic DNA sample to be identified specifically comprises: Using the genomic DNA of the mouse to be identified as a template, multiplex PCR amplification is performed using an amplification primer combination for amplifying the 32 SNP sites to obtain an amplified product; Using SAP enzyme to perform enzymatic digestion reaction on the amplified product to obtain an enzymatic digestion product; Using the single-base extension composition of the 32 SNP sites to extend the enzymatic digestion product to obtain an extension product; The extension products were analyzed by mass spectrometry using MALDI-TOF, and the genotype of the SNP site combination was obtained based on the analysis results.

10. A method for detecting or assisting in detecting the genetic quality of mice, characterized in that: include: Obtain genomic DNA of the mouse to be tested; Detecting the genotype of the SNP site combination according to claim 1 in the mouse genomic DNA sample to be identified; The genotype of the SNP site combination of the mouse to be identified is compared with the standard genotype of the SNP site combination of the target strain mouse. When the genotype of the SNP site combination of the mouse to be identified is the same as the standard genotype of the SNP site combination of the target strain mouse, the genetic quality is judged to be stable, otherwise the genetic quality is unqualified.

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