Combinations of snp loci and methods for identifying or aiding in the identification of a mouse genetic background

Abstract

The application provides a SNP site combination and a method for identifying or assisting in identifying a mouse genetic background. The application provides a SNP site combination for identifying or assisting in identifying a mouse genetic background, which comprises 32 SNP sites. The application provides a method for identifying or assisting in identifying a mouse genetic background based on the SNP site combination, which realizes identification or assistance in identifying the genetic background of a mouse to be identified by matching the genotype of the SNP site combination of the mouse to be identified with a reference gene profile of a SNP site combination of a target strain mouse. The method provided by the application has the characteristics of high cost performance, fast 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 application relates to the field of biotechnology, and particularly relates to a SNP site combination for identifying or assisting in identifying the genetic background of mice and a method. BACKGROUND

[0002] Genetic quality monitoring is an important part of experimental animal quality control, and a clear genetic background is a prerequisite for ensuring the reliability and accuracy of scientific research. Traditional genetic background detection methods for experimental animals include immunological 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 been widely used, and SNP, as the third generation of genetic markers, has the advantages of large number, wide distribution, high genetic type, strong representation and easy analysis. Providing a SNP site combination for identifying the genetic background of mice is a technical problem to be solved in the art. SUMMARY

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

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

[0005] Table 1: 32 SNP site combination

[0006]

[0007]

[0008] In Table 1, the positions of rs3022953, rs3023442 and rs3023450 sites use GRCm38 version, and the positions of the remaining sites use GRCm39 version.

[0009] The application as described above, the substance for detecting the genotype of the SNP site combination comprises an amplification primer composition for amplifying 32 SNP sites and a single base extension composition; the amplification primer composition comprises single-stranded DNA molecules with nucleotide sequences of SEQ ID NO: 1-64; and the single base extension composition comprises single-stranded DNA molecules with nucleotide sequences of SEQ ID NO: 65-96.

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

[0011] In a third aspect, the present application provides a single base extension composition for detecting the genotype of the SNP site combination, comprising single-stranded DNA molecules with nucleotide sequences of SEQ ID NO: 65-96.

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

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

[0014] The product as described above can further comprise other reagents required for detecting the genotype of the SNP site combination, for example, can comprise DNA polymerase, dNTP, etc. required for PCR, and can further comprise SAP enzyme (Shrimp Alkaline Phosphatase), etc.

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

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

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

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

[0019] Taking the genotype of the SNP site combination of a target strain of mice as a standard gene profile, comparing the genotype of the SNP site combination of the mouse to be identified with the standard gene profile, and identifying or assisting in identifying the genetic background of the mouse to be identified.

[0020] The mouse strain as described above refers to a population consisting of a group of mice with similar genetic characteristics in a specific genetic background. Different mouse strains have different genetic backgrounds. The target strain refers to a mouse with a known genetic background and a determined strain.

[0021] The method as described above, the mouse is one of inbred line, congenic line or hybrid population, which can be determined according to the experimental requirements; in the present application, the inbred mouse may, for example, be at least one of 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB, SJL; the congenic mouse may, for example, be at least one of SCID-Beige, B6 Albino, APOE, CB-17SCID; the hybrid mouse may, for example, be at least one of CB6F1 and B6D2F1.

[0022] The method as described above, the mouse strain is defined according to the People's Republic of China National Standard numbered GB-14923-2022, specifically, the 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; the congenic line refers to a line that is only different from the original inbred line in a very small chromosomal fragment by backcrossing a specific marker from the donor strain; the hybrid population refers to the offspring population produced by crossing two different inbred lines.

[0023] The method as described above, the mouse to be identified can be one of inbred line, congenic line or hybrid population. When the mouse to be identified is suspected to be an inbred line or a hybrid population, the corresponding target strain mouse selects the same inbred line or hybrid population. 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 suspected to be a congenic line, if the genotypes of the above SNP sites of the mouse to be identified and the target strain mouse are the same, it is inferred that the mouse to be identified is derived from the genetic background of the target strain mouse.

[0024] The method as described above, the genotype of the SNP site combination of the target strain mouse can be obtained from the database, or can be obtained by testing the SNP site combination genotype of the known strain mouse; wherein the database can be National Center for Biotechnology Information (NCBI) or Mouse PhenomeDatebase.

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

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

[0027] An enzymatic reaction is performed on the amplification product using SAP enzyme to obtain an enzymatic product.

[0028] The single base extension composition of 32 SNP sites is used to extend the enzymatic product to obtain an extension product;

[0029] The extension product is subjected to mass spectrometry analysis using MALDI-TOF, and the genotype of the SNP site combination is obtained according to the analysis result.

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

[0031] Obtaining the genomic DNA of a mouse to be detected;

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

[0033] Comparing the genotype of the SNP site combination of the mouse to be identified with the standard genotype of the SNP site combination of the target strain mouse, and 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, it is determined that the genetic quality is stable, otherwise the genetic quality is unqualified.

[0034] The present application provides a SNP site combination for identifying or assisting in identifying the genetic background of a mouse, and provides a method for identifying or assisting in identifying the genetic background of a mouse based on the SNP site combination. According to the method provided by the present application, 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 application 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 DRAWINGS

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

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

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

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

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

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

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

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

[0043] Figure 9 MALDI-TOF mass spectrum of the mouse rs3023381 locus for the mouse numbered 20240910-6 (C57BL / 6N);

[0044] Figure 10 MALDI-TOF mass spectrum of the mouse rs3023382 locus for the mouse numbered 20240910-6 (C57BL / 6N);

[0045] Figure 11 MALDI-TOF mass spectrum of the mouse rs3023442 locus for the mouse numbered 20240910-6 (C57BL / 6N);

[0046] Figure 12 MALDI-TOF mass spectrum of the mouse rs3023481 locus for the mouse numbered 20240910-6 (C57BL / 6N);

[0047] Figure 13 MALDI-TOF mass spectrum of the mouse rs3088673 locus for the mouse numbered 20240910-6 (C57BL / 6N);

[0048] Figure 14 MALDI-TOF mass spectrum of the mouse rs3089604 locus for the mouse numbered 20240910-6 (C57BL / 6N);

[0049] Figure 15MALDI-TOF mass spectrum of the mouse rs3091174 locus numbered 20240910-6 (C57BL / 6N);

[0050] Figure 16 MALDI-TOF mass spectrum of the mouse rs3659787 locus numbered 20240910-6 (C57BL / 6N);

[0051] Figure 17 MALDI-TOF mass spectrum of the mouse rs3702158 locus numbered 20240910-6 (C57BL / 6N);

[0052] Figure 18 MALDI-TOF mass spectrum of the mouse rs3706082 locus numbered 20240910-6 (C57BL / 6N);

[0053] Figure 19 MALDI-TOF mass spectrum of the mouse rs3709624 locus numbered 20240910-6 (C57BL / 6N);

[0054] Figure 20 MALDI-TOF mass spectrum of the mouse rs3722313 locus numbered 20240910-6 (C57BL / 6N);

[0055] Figure 21 MALDI-TOF mass spectrum of the mouse rs13481123 locus numbered 20240910-6 (C57BL / 6N);

[0056] Figure 22 MALDI-TOF mass spectrum of the mouse rs13481154 locus numbered 20240910-6 (C57BL / 6N);

[0057] Figure 23 MALDI-TOF mass spectrum of the mouse rs3022796 locus numbered 20240910-6 (C57BL / 6N);

[0058] Figure 24 MALDI-TOF mass spectrum of the mouse rs3022825 locus numbered 20240910-6 (C57BL / 6N);

[0059] Figure 25 MALDI-TOF mass spectrum of the mouse rs3022953 locus numbered 20240910-6 (C57BL / 6N);

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

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

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

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

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

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

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

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

[0068] In order to make the objects, technical solutions and advantages of the present application clearer, the technical solutions in the present application will be described clearly and completely below in conjunction with the accompanying drawings in the present application. Obviously, the described embodiments are some embodiments of the present application, rather than all the embodiments. They should not be understood as limiting the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application. In the description of the present application, it should be understood that the terms used are only for the purpose of description, and should not be understood as indicating or implying relative importance.

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

[0070] Example 1

[0071] 1. Combinations of 32 SNP sites and substances for detecting the combinations of 32 SNP sites

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

[0073] Table 2 Combinations of 32 SNP sites and primers for detecting the SNP sites

[0074]

[0075]

[0076]

[0077] 2. Establishment of a SNP site combination genotyping 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 individual amplification primers were re-dissolved in deionized water to obtain a storage solution with a concentration of 100 μmol / L. Equal volumes of the storage solutions of the 64 amplification primers corresponding to the 32 SNP site combinations were mixed, deionized water was added to make up the volume, and an amplification primer composition with a final concentration of 0.5 μmol / L for each amplification primer was obtained, which was designated as 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) was calculated as (target reaction concentration * 9) / 0.94 * extension primer mixing number (e.g. 15 extension primers mixed, the number is 15), and the amount of deionized water (μL) required to be added to each primer tube was calculated as extension primer OD * 33 / extension primer Mass / target working concentration * 1000000. The primers were re-dissolved in the calculated amount to obtain an extension primer storage solution. Equal volumes of the extension primer storage solutions were mixed to obtain a single-base extension composition, which was designated as extension primer Mix.

[0081] 2.2. Testing of the genotypes of SNP site combinations of 16 common strain mice

[0082] 2.2.1, Collect 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB and SJL 10 inbred strains of mice, CB6F1 and B6D2F1 2 hybrid population of mice, SCID-Beige, B6 Albino, APOE and CB-17 SCID 4 of the same type of mice, each strain randomly selected 3. A total of 48 mice tail, tail length is not less than 0.5 cm.

[0083] 2.2.2, using the automatic nucleic acid extractor, using the ABI magnetic bead method nucleic acid extraction kit from the mouse tail to extract genomic DNA, get the concentration of 10-20 ng / μL of template DNA solution. 20 μL of template DNA solution to 96-well plate arranged in turn, do a good job of labeling for use.

[0084] 2.2.3, using Agena company supporting PCR reagent, according to the PCR reaction system shown in table 3 to prepare 10% of the amount of reagent for detection, the prepared mixture with the volume of 4 μL / well into the PCR plate hole, finally add 1 μL / well of template DNA, get the total volume of 5 μL / well of PCR reaction system. According to the PCR reaction program shown in table 4, PCR amplification, get the amplification product.

[0085] Table 3 PCR reaction system

[0086] Component Final concentration Volume (μL) Water / 1.8 Amplification primer Mix (0.5 μM) 0.1 μM 1 10*Buffer (with Mg 2+ )]]> 2 mM MgCl2 0.5 MgCl2(25 mM) 2 mM 0.4 dNTP (25 mM) 500 μM 0.1 PCR enzyme 1 Unit 0.2 Template DNA (10 ng / μL) 1 Total amount 5

[0087] Table 4 PCR reaction program

[0088]

[0089] 2.2.4, enzymatic reaction

[0090] using Agena iPLEX Gold kit, according to the SAP reaction system shown in table 5, according to the amount of 10% of the detection of the preparation of the mixture, named SAP enzyme Mix, in 2.2.3 obtained in each amplification product, add 2 μL / well of prepared SAP enzyme Mix, blow and mix, according to the enzymatic reaction program shown in table 6 in the PCR instrument SAP enzymolysis reaction, get the enzymolysis product.

[0091] Table 5 SAP reaction system

[0092] Component Volume (μL) SAP buffer 0.17 SAP enzyme 0.3 Water 1.53 Total amount 2

[0093] Table 6 SAP enzymolysis reaction program

[0094]

[0095]

[0096] 2.2.5, Single base extension reaction

[0097] Using Agena iPLEX Gold kit, the reaction system shown in Table 7 was prepared by 10% more detection amount of mixture, named single base extension reaction Mix, 2 μL / well of single base extension reaction Mix prepared in the above enzymatic product was added, and the mixture was mixed by blowing and sucking. The single extension reaction was carried out in PCR instrument according to the reaction program shown in Table 8 to obtain the extension product.

[0098] Table 7 Single base extension reaction system

[0099] Component Volume (μL) UEP primer Mix 0.94 Gold buffer 0.2 Stop buffer 0.1 iPlex enzyme 0.04 Water 0.62 Total amount 2

[0100] Table 8 Single base extension reaction program

[0101]

[0102] 2.2.6, Mass spectrometry detection

[0103] The extension product was desalted and purified, and the purified product was transferred to the chip corresponding matrix point using MassARRAY Nanodispenser RS1000 equipment. MassARRAY Analyzer was used for detection, and the detection genotypes of 32 SNP sites of 48 mouse samples were obtained. The detection results are shown in Table 9. The MALDI-TOF mass spectrum of 32 SNP sites of the mouse numbered (sample Id) 20240910-6 (C57BL / 6N) is shown in Figures 1-32 .

[0104] The detection of 48 samples showed that 9 sites showed no call, respectively, rs3023064 site of sample 20240910-48, rs3089604 and rs3722313 sites of sample 20240910-22, rs3089604, rs3722313 and rs3023064 sites of sample 20240910-38, rs3022796, rs3022977 and rs13481123 of sample 20240910-21, and the genotypes shown in Table 9 were manually read according to the mass spectrum peak. (When the no call result is concentrated in a certain sample and other samples are normal, it is considered that the problem of nucleic acid quality of the sample causes.) The lowest call rate of the sample was 90.6%, and the highest was 100%. All meet more than 90%, and the detection rate is good.

[0105] According to Table 9, the genotypes of 32 SNP sites detected by the method provided in Table 9 are compared with the reference gene profile of the mouse, and it is found that the detection results shown in Table 9 are 100% matched with the actual situation, indicating that the accuracy of the method provided in the application is high. The genotypes of 32 SNP sites of 16 strain mice provided in the application can be distinguished from each other, indicating good specificity. The genotypes of 32 SNP sites of 3 different samples of the same strain provided in the application are consistent (except BALB / c Nude), indicating good stability.

[0106] 3. Comparison of SNP site genotypes of 16 strain mice with reference gene profile

[0107] In the database Mouse Phenome Database, the genotypes of 32 SNP sites of 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 II2rg tm1WjI / SzJ (NOG), NOD / Shi Lt J (NOD SCID), BALB / c nude strain mice were queried, and the query results are shown in Table 10 as a reference gene profile. The reference gene profile of the hybrid group mice CB6F1, B6D2F1 is obtained by detecting the standard genotype of the parent strain.

[0108] The SNP site combination genotypes of the 16 strains shown in Table 9 are compared with the reference gene map shown in Table 10, if the SNP sites of the 16 strains of mice are consistent with the genotype of the reference gene map, the number of gene matches is recorded as 1, otherwise as 0, the value corresponding to 32 SNP sites of each sample is calculated = gene matching number / 32*100%, and the calculation result is shown in Table 11. According to Table 11, the detection results of inbred and hybrid group mice are 100% matched with the standard gene profile of the corresponding strain mice, indicating that the method provided by the application can distinguish and identify these inbred and hybrid group mice; the sample detection results of SCID-Beige and CB-17SCID strains are 100% matched with the BALB / cJ reference gene profile, the sample detection results of B6 Albino are 100% matched with the C57BL / 6NJ reference gene profile, and the sample detection results of APOE are 100% matched with the C57BL / 6J reference gene profile, indicating that the same strain strains cannot be successfully distinguished, but the background sources of these same strain strains can be clearly determined.

[0109] Finally, it should be noted that: the above examples are only used to illustrate the technical solutions of the present application, and not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that they can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

Claims

1. Use of a substance for detecting the genotype of a combination of SNP loci in the identification or aid in the identification of the genetic background of mice, characterized in that, The SNP site combination comprises 32 SNP sites shown in Table 1: the mouse is selected from at least one of 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB, SJL, CB6F1, B6D2F1, SCID-Beige, B6 Albino, APOE and CB-17 SCID; Table 1 32 SNP site combination In Table 1, the positions of rs3022953, rs3023442 and rs3023450 sites are in the version of GRCm38, and the positions of the remaining sites are in the version of GRCm39.

2. Use according to claim 1, characterized in that, The substance for detecting the genotype of the SNP site combination comprises an amplification primer composition and a single base extension composition for amplifying 32 SNP sites; the amplification primer composition comprises single-stranded DNA molecules with the nucleotide sequences of SEQ ID NOs: 1-64; and the single base extension composition comprises single-stranded DNA molecules with the nucleotide sequences of SEQ ID NOs: 65-96.

3. A product for detecting the genotype of the combination of SNP loci according to claim 1, characterized in that, The product comprises an amplification primer composition and a single base extension composition; wherein the amplification primer composition comprises single-stranded DNA molecules with the nucleotide sequences of SEQ ID NOs: 1-64, and the single base extension composition comprises single-stranded DNA molecules with the nucleotide sequences of SEQ ID NOs: 65-96.

4. The product of claim 3 for use in identifying or assisting in identifying the genetic background of a mouse selected from at least one of 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB, SJL, CB6F1, B6D2F1, SCID-Beige, B6 Albino, APOE and CB-17 SCID.

5. A method of identifying or aiding in the identification of a mouse genetic background, characterized in that, The method comprises: obtaining genomic DNA of a mouse to be identified; detecting the genotype of the SNP site combination of claim 1 in the genomic DNA sample of the mouse to be identified; comparing the genotype of the SNP site combination of the mouse to be identified with a standard gene profile using the genotype of the SNP site combination of a target strain mouse as the standard gene profile, to identify or assist in identifying the genetic background of the mouse to be identified; the mouse is at least one of an inbred strain, a congenic strain or a hybrid population, the mouse of the inbred strain is at least one of 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB and SJL, the mouse of the hybrid population is at least one of CB6F1 and B6D2F1, and the mouse of the congenic strain is at least one of SCID-Beige, B6 Albino, APOE and CB-17 SCID.

6. The method of claim 5, wherein, Detecting the genotype of the SNP site combination of claim 1 in the genomic DNA sample of the mouse to be identified, specifically comprising: Using the genomic DNA of the mouse to be identified as a template, using the amplification primer composition for amplifying 32 SNP sites to perform multiplex PCR amplification to obtain an amplification product; Using SAP enzyme to perform enzymatic reaction on the amplification product to obtain an enzymatic product; Using the single base extension composition of 32 SNP sites to perform extension on the enzymatic product to obtain an extension product; Using MALDI-TOF to perform mass spectrometry analysis on the extension product, and obtaining the genotype of the SNP site combination according to the analysis result.

7. A method of detecting or aiding in the detection of genetic quality in mice, characterized in that, Comprising: Obtaining the genomic DNA of the mouse to be detected; Detecting the genotype of the SNP site combination of claim 1 in the genomic DNA sample of the mouse to be identified; Comparing the genotype of the SNP site combination of the mouse to be identified 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, it is judged that the genetic quality is stable, otherwise the genetic quality is unqualified; The mouse is at least one of an inbred strain, a homologous strain or a hybrid group, the mouse of the inbred strain is at least one of 129, BALB / c, BALB / c Nude, C3H, C57BL / 6JNifdc, C57BL / 6N, DBA / 1, DBA / 2, FVB and SJL, the mouse of the hybrid group is at least one of CB6F1 and B6D2F1, and the mouse of the homologous strain is at least one of SCID-Beige, B6 Albino, APOE and CB-17 SCID.

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