Method for detecting SNP of ADK gene of Anhui white goat and breeding application
By detecting the SNP site of the ADK gene in Anhui white goats using PCR amplification and Sanger sequencing, the problems of complexity and high cost in existing technologies have been solved, achieving simple, low-cost, and highly accurate detection, thus improving breeding efficiency.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- ANHUI AGRICULTURAL UNIVERSITY
- Filing Date
- 2026-03-24
- Publication Date
- 2026-05-12
AI Technical Summary
Existing SNP detection methods cannot achieve specific, low-cost, and high-accuracy detection of the seventh intron of the ADK gene in Anhui white goats, making it difficult to meet the needs of molecular marker-assisted breeding.
Using PCR amplification combined with agarose gel electrophoresis and Sanger sequencing, specific primers were designed to amplify and detect the seventh intron of the ADK gene in Anhui white goats, identifying three SNP sites. Individuals with excellent growth traits were selected by genotyping rules.
This method enables simple, low-cost, and highly accurate detection of ADK gene SNPs, clarifying the significant correlation between SNP sites and growth traits, and improving breeding efficiency.
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Figure CN122012744A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of molecular genetic breeding technology, specifically to a method for detecting SNPs in the ADK gene of Anhui white goats and its breeding applications. Background Technology
[0002] Molecular marker-assisted selection is a core technology in modern livestock and poultry breeding. It utilizes DNA molecular markers closely linked to target traits to achieve precise genotypic selection in the early stages of breeding, shortening the breeding cycle and improving the efficiency of genetic improvement. Single nucleotide polymorphisms (SNPs) are genetic markers formed by single nucleotide variations in the genome. They can influence important economic traits in livestock and poultry by regulating gene expression and protein function, and are key markers in molecular breeding.
[0003] Current SNP detection methods include Sanger sequencing, pyrosequencing, high-resolution melting curve analysis, PCR-LDR, allele-specific PCR, TaqMan probe method, and SNaPshot method. However, pyrosequencing requires specialized equipment, high-resolution melting curve analysis demands high instrument stability, PCR-LDR has a long preparation period, TaqMan probe method has low throughput, and SNaPshot method is costly; none of these methods can simultaneously achieve simplicity, low cost, and high accuracy.
[0004] Adenosine kinase (ADK) genes are widely involved in adenosine metabolism and epigenetic regulation. In humans and model animals, they have been shown to be associated with angiogenesis, muscle regeneration, and nervous system diseases, but research in goats is scarce. Anhui White Goat, an important local meat goat breed, lacks ADK gene SNP molecular markers that are directly related to growth traits. Current technologies cannot achieve specific, low-cost, and high-accuracy SNP detection for the seventh intron of the ADK gene in this breed, making it difficult to meet the needs of marker-assisted breeding. Summary of the Invention
[0005] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a method and application for detecting intron 7 single nucleotide polymorphisms in the ADK gene of Anhui white goats that is simple to operate, low in cost, and highly accurate, thereby solving the problems of complex existing SNP detection methods, lack of functional markers for the ADK gene in Anhui white goats, and low breeding efficiency.
[0006] To achieve the above objectives, the present invention provides the following technical solution:
[0007] This invention proposes a method for detecting SNPs in the ADK gene of Anhui white goats. The method involves using the whole-genome DNA of the Anhui white goat to be tested as a template, amplifying the seventh intron fragment of the ADK gene via PCR, and performing Sanger sequencing after confirming the correct band by agarose gel electrophoresis. Three SNP sites were identified: NC_030835.1:g.15498308, NC_030835.1:g.15498501, and NC_030835.1:g.15498816.
[0008] The primers used for the PCR amplification are:
[0009] Upstream primer: 5'-TGATGACCTGCATTAAGCACCA-3';
[0010] Downstream primer: 5'-AGCCATAATCCGATCTGAGGTAA-3'.
[0011] Preferably, the PCR amplification reaction program is as follows: pre-denaturation at 93-95 ℃ for 3-5 min; denaturation at 95-98 ℃ for 10-30 s, annealing at 55-60 ℃ for 30-50 s, extension at 65-72 ℃ for 1-2 min, for 30-35 cycles; extension at 65-72 ℃ for 3-5 min, and storage at 4 ℃.
[0012] Preferably, the PCR amplification reaction system includes: 12.5 μL of 2×PCR MasterMix, 25–50 ng of whole genome DNA of Anhui white goats to be tested, and 2.5–5 pmol each of upstream and downstream primers for amplification, with ultrapure water added to bring the total volume to 25 μL.
[0013] Preferably, the agarose gel electrophoresis uses an agarose gel with a mass fraction of 1.5% to 2.0%, and the PCR amplification product fragment size is 767 bp.
[0014] Preferably, the genotype determination rule for the Sanger sequencing is as follows:
[0015] The NC_030835.1:g.15498308 locus has the genotype AA when it is a single peak and the genotype AG when it is a multi-peaked locus.
[0016] At the NC_030835.1:g.15498501 locus, the genotype is CC or GG when there is a single peak, and CG when there are multiple peaks.
[0017] The NC_030835.1:g.15498816 locus has a genotype of TT or GG when it is a single peak and a genotype of TG when it is a multi-peak.
[0018] Preferably, a method for molecular marker-assisted selection breeding of Anhui white goats involves detecting the SNPs of the ADK gene in Anhui white goats, detecting the genotypes of three SNP loci in the intron 7 of the ADK gene in Anhui white goats, and using the three SNP loci as molecular markers to screen individuals with excellent growth traits.
[0019] Preferably, the growth traits include body weight, body height, body length, chest circumference, tube circumference, and rump width.
[0020] Preferably, individuals of Anhui white goats with the AA genotype at NC_030835.1:g.15498308, the GG genotype at NC_030835.1:g.15498501, and the GG genotype at NC_030835.1:g.15498816 are selected as the core breeding group.
[0021] The beneficial effects of this invention are as follows:
[0022] (1) This invention designs primers for PCR amplification of three SNP sites on the seventh intron of the ADK gene. The genotype of an individual can be determined by PCR, agarose gel electrophoresis detection and Sanger sequencing analysis. No special expensive equipment is required. The operation is simple, the cost is low and the results are accurate, which solves the defects of existing SNP detection methods that are heavily dependent on equipment and complicated to operate.
[0023] (2) This invention identifies three SNP sites in the seventh intron of the ADK gene of Anhui white goat for the first time. Among them, SNP1 is a newly discovered site. It provides dedicated specific primers, and the amplified fragment is stable at 767bp, with strong detection specificity.
[0024] (3) This invention clearly identifies three SNP loci that are significantly associated with the weight, height, length, chest circumference, cannon circumference, and rump width of Anhui white goats, providing a combination of advantageous genotypes that can be directly used for molecular marker-assisted selection, which is beneficial to accelerating the breeding process of superior goat breeds. Attached Figure Description
[0025] Figure 1 This is an electrophoresis diagram of the amplified sequence of the seventh intron of the ADK gene of Anhui white goat in an embodiment of the present invention.
[0026] Figure 2 This is a sequencing peak diagram of the AA and AG genotypes at the SNP1 site on the seventh intron sequence of the goat ADK gene in an embodiment of the present invention.
[0027] Figure 3 This is a sequencing peak diagram of the CC, GG, and CG genotypes at the SNP2 site on the seventh intron sequence of the goat ADK gene in an embodiment of the present invention.
[0028] Figure 4 This is a sequencing peak diagram of the TT, GG, and TG genotypes at the SNP3 site on the seventh intron sequence of the goat ADK gene in an embodiment of the present invention. Detailed Implementation
[0029] The present invention will be described in further detail below with reference to specific preparation and application examples and data. It should be understood that these examples are merely illustrative of the invention and are not intended to limit the scope of the invention in any way.
[0030] In the following embodiments, various processes and methods not described in detail are conventional methods known in the art. All primers used are indicated upon their first appearance, and subsequent use of the same primer will use the same indication as the initial indication.
[0031] Example 1:
[0032] The specific steps for screening the seventh intron SNP site of the ADK gene in Anhui white goats are as follows:
[0033] Searching for SNP sites in the goat ADK gene in the Ensembl database (http: / / lasia.ensembl.org / index.html) revealed two SNP mutation sites in the seventh intron region of the ADK gene of Anhui white goats (rs670723959,NC_030835.1:g. 15498501, C>G; rs656652252,NC_030835.1:g. 15498816, T>G).
[0034] Using the goat ADK gene (Gene ID: 102189651) as a reference sequence, primers were designed using NCBI primer Blast (https: / / www.ncbi.nlm) to amplify DNA sequences containing the above variant sites targeting the above SNP sites.
[0035] The whole genome DNA of 30 Anhui white goats was mixed and used as a PCR amplification template. The amplified products were subjected to agarose gel electrophoresis at a concentration of 1.5-2.0% (w / v). The size of the electrophoretic bands was used to preliminarily determine whether the amplified bands were correct. If the size was basically in line with expectations, the PCR amplification products were sent to a sequencing company for Sanger sequencing to identify the genetic variation sites in the seventh intron region of the ADK gene. Subsequently, according to the sequencing peak diagram, it was determined that there were 3 SNP mutation sites in the seventh intron region of the amplified Anhui white goat ADK gene (NC_030835.1:g.15498308, A>G; rs670723959, NC_030835.1:g.15498501, C>G; rs656652252, NC_030835.1:g.15498816, T>G).
[0036] Example 2:
[0037] The specific steps for PCR amplification and electrophoresis detection are as follows:
[0038] 1. Preparation of experimental reagents and solutions:
[0039] 2×Magic Green Taq Super Mix (TOLOBIO, Shanghai); Animal Tissue / Cell Genomic DNA Extraction Kit (Adley, Beijing); DNA Marker DL2000 (Qingke Biotechnology, Beijing); Agarose (Spain); Nucleic Acid Dyes (Adley, Beijing); 50×TAE (Solepro, Beijing)
[0040] All reaction systems, solutions, and buffer solutions were prepared using deionized ultrapure water and autoclaved at 15 bf / in (1.034 × 10⁻⁶). 5 Pa), 25 min. The specific dilution method for 1×TAE solution is as follows: Add 10 mL of 50×TAE to 490 mL of deionized ultrapure water to prepare 500 mL of 1×TAE buffer solution, and store at room temperature for later use.
[0041] 2. Design amplification primers targeting candidate DNA regions:
[0042] Using the goat ADK gene sequence (Gene ID: 102189651) published in the NCBI database (http: / / www.ncbi.nlm.nih.gov / ) as a primer design template, amplification primers containing the seventh intron region of the goat ADK gene were designed using the Primer-BLAST website to amplify the region containing the polymorphic site. The designed primers amplified a target fragment of 767 bp (primer pair P1).
[0043] Upstream primer F1: 5'-TGATGACCTGCATTAAGCACCA-3';
[0044] Downstream primer R1: 5'-AGCCATAATCCGATCTGAGGTAA-3'.
[0045] 3. PCR amplification to detect individual genotype:
[0046] The PCR reaction system uses a mixed loading method. Based on the amount of each component required for each reaction system and the number of PCR reactions required for one reaction, the total amount of each component is calculated and added to a 1.5 mL centrifuge tube. After thorough mixing, the tube is briefly centrifuged and then aliquoted into each 0.2 mL Eppendorf PCR tube. Template DNA (goat genomic DNA) is then added, followed by brief centrifugation and PCR amplification.
[0047] The PCR reaction system consisted of: 12.5 μL of 2×Magic Green Taq Super Mix (containing Taq DNA polymerase, dNTPs, and reaction buffer); 0.5 μL of upstream primer (10 μmol / L); 0.5 μL of downstream primer (10 μmol / L); 0.5 μL of genomic DNA (concentration of 50 ng / μL genomic DNA); and 11.0 μL of deionized water; for a total of 25.0 μL.
[0048] The PCR reaction procedure was as follows: pre-denaturation at 93–95 ℃ for 3–5 min; denaturation at 95–98 ℃ for 10–30 s, annealing at 55–60 ℃ for 30–50 s, extension at 65–72 ℃ for 1–2 min, for 30–35 cycles; extension at 65–72 ℃ for 3–5 min, and storage at 4 ℃.
[0049] 4. Preparation of 1.5–2.0% agarose gel and electrophoretic detection:
[0050] 1) Clean and dry the agarose gel box and plate. Place the cleaned gel preparation base plate into the gel box and insert the comb.
[0051] 2) The agarose gel concentration is the ratio of the mass (g) of agarose to the volume (mL) of 1×TAE buffer. For example, to prepare a 2.0% agarose gel, weigh 2.0 g of agarose, transfer it to an Erlenmeyer flask, add 100 mL of 1×TAE buffer, shake to dissolve, and use a heating device such as a microwave oven to fully dissolve the agarose. After boiling twice, remove the flask and let it cool until it is no longer hot to the touch, then add 5 μL of nucleic acid dye. Gently shake to mix, preventing air bubbles from forming.
[0052] 3) After mixing, immediately pour the agarose solution into the gel box; if air bubbles appear, immediately remove them with a pipette.
[0053] 4) After the gel has completely cooled and solidified (about 25 to 40 minutes), remove the comb and transfer the gel into the electrophoresis tank.
[0054] 5) Add 1×TAE buffer to the electrophoresis tank so that the liquid level is 2-5 mm above the gel level.
[0055] 6) Add 3-5 μL of DNA sample to the wells and add DNA Marker to one or both wells.
[0056] 7) Electrophoresis at 110 V to 220 V for 25 to 30 minutes.
[0057] 8) Imaging was performed using the Bio-Rad gel imaging system, and fragment sizes were analyzed based on the images. The goat genome was then amplified by PCR using the primers described above. The electrophoretic detection results of the amplification products are as follows: Figure 1 As shown, the marker lanes are Marker D2000 (from largest to smallest: 2000 bp, 1000 bp, 750 bp, 500 bp, 250 bp, 100 bp). Lanes 1-9 represent the target fragment (767 bp) amplified from different samples.
[0058] Example 3:
[0059] The specific steps for Sanger sequencing and genotyping are as follows:
[0060] 1. Send PCR products with the correct band size from agarose gel analysis to a sequencing company for Sanger sequencing.
[0061] 2. Genotype Determination: Genotype determination was performed based on the sequencing results returned by the sequencing company. A single peak indicates a homozygous genotype, while overlapping peaks indicate a heterozygous genotype. Results are as follows: Figure 2 , Figure 3 and Figure 4 As shown.
[0062] SNP1 (NC_030835.1:g.15498308): Figure 2 The sequence results for individuals with different genotypes at the SNP1 locus are shown. When the red arrow indicates a locus with a single green peak, the individual has the AA genotype. When the red arrow indicates a locus with overlapping green and red peaks, the individual has the AG genotype.
[0063] SNP2 (NC_030835.1:g.15498501): Figure 3The sequence results for individuals with different genotypes at the SNP2 locus are shown. When the red arrow indicates a locus with a single blue peak, the individual has the CC genotype; when the red arrow indicates a locus with a single black peak, the individual has the GG genotype; and when the red arrow indicates a locus with overlapping blue and black peaks, the individual has the CG genotype.
[0064] SNP3 (NC_030835.1:g.15498816): Figure 4 The sequence results for individuals with different genotypes at the SNP3 locus are shown. When the red arrow indicates a single red peak, the individual has the TT genotype; when the red arrow indicates a single black peak, the individual has the GG genotype; and when the red arrow indicates a combination of red and black peaks, the individual has the TG genotype.
[0065] The above results indicate that by using the designed primers to amplify the goat ADK genomic DNA by PCR and interpreting the genotype using Sanger sequencing, it is possible to rapidly and accurately identify the three SNP sites in the seventh intron of the goat ADK gene.
[0066] Example 4:
[0067] The specific steps for statistical analysis of genetic variation frequencies are as follows:
[0068] Using 180 Anhui white goats as the test group, genotype frequencies and allele frequencies were statistically analyzed, and Hardy-Weinberg equilibrium tests were performed.
[0069] 1. Gene and genotype frequency analysis:
[0070] Genotype frequency refers to the ratio of individuals with a certain genotype to the total number of individuals in a population. The calculation formula is as follows:
[0071] P YY =N YY / N,
[0072] In the formula, P YY The frequency of the YY genotype at a specific locus; N YY This represents the number of individuals in the population with the YY genotype; N is the total number of individuals in the population being tested.
[0073] Gene frequency refers to the ratio of the number of a particular gene to the total number of its alleles in a population. The calculation formula is as follows:
[0074] P Y =(2N Y +N Ya1 +N Ya2 +N Ya3 +N Ya4 + ... + N Yan ) / 2N,
[0075] In the formula, P Y N represents the frequency of the Y allele. Y N represents the number of individuals in the population with the Y genotype. Yai Indicates that the group has Ya i (i=1,2,…n) Number of individuals with genotypes, a1 to a n Let Y be a set of n distinct multiple alleles.
[0076] 2. Results Analysis:
[0077] Table 1 shows the allele frequencies and genotype frequencies of the three SNP loci on the seventh intron of the ADK gene in Anhui white goats.
[0078] Table 1. Frequency distribution of three SNP sites in the ADK gene of Anhui white goats.
[0079]
[0080] (1) SNP1 (NC_030835.1:g.15498308) locus: There are two genotypes at this locus, AA and AG, with frequencies of 0.92 and 0.02, respectively. The frequencies of the A and G alleles are 0.93 and 0.07, respectively. Hardy-Weinberg equilibrium analysis showed that this locus is in Hardy-Weinberg equilibrium.
[0081] (2) SNP2 (NC_030835.1:g.15498501) locus: There are three genotypes at this locus, CC, CG and GG, with frequencies of 0.82, 0.15 and 0.03, respectively. The frequencies of C and G alleles are 0.90 and 0.11, respectively. Hardy-Weinberg equilibrium analysis found that this locus is in Hardy-Weinberg disequilibrium.
[0082] (3) SNP3 (NC_030835.1:g.15498816) locus: There are 3 genotypes at this locus, TT, TG and GG, with frequencies of 0.83, 0.16 and 0.02, respectively. The frequencies of T and G alleles are 0.90 and 0.10, respectively. Hardy-Weinberg equilibrium analysis showed that this locus is in Hardy-Weinberg equilibrium.
[0083] The above results indicate that all three SNP loci exhibit genetic polymorphism in the Anhui white goat population and can be used as molecular markers for trait association analysis.
[0084] Example 5:
[0085] The specific steps for SNP site-growth trait association analysis are as follows:
[0086] One hundred and eighty 36-month-old female Anhui white goats were used as the experimental group. Under consistent feeding and management conditions, body weight, height, length, chest circumference, cannon bone circumference, and rump width were measured. SPSS 25.0 software was used to analyze the correlation between breed, environment, and gene loci and growth traits.
[0087] First, descriptive statistical analysis is performed on the obtained data to determine if outliers exist. Then, based on the characteristics of the data, analysis of variance (when three genotypes exist, and the number of individuals with each genotype is greater than or equal to 3), multiple linear models, or t-analysis (when only two genotypes exist, and the number of individuals with each genotype is greater than or equal to 3) are used to analyze the effect of genotype. During data processing, the effects of environment, breed, age, sex, and genotype are considered according to the different factors affecting the indicators of growth traits. However, since all samples come from the same farm, with consistent feeding methods, and the individuals are all 36-month-old adult female Anhui White Goats, environment, breed, age, and sex can be considered as fixed factors, and the statistical analysis model can be simplified to the following statistical model:
[0088] Y ij =μ+Genotype i +e j
[0089] In the formula, Y ij Individual phenotypic record; μ: population mean; Genotype i : Genotype effect; e ij Random error.
[0090] Association analysis showed that all three detected SNP loci were significantly associated with body size traits in Anhui white goats. Specific results are as follows:
[0091] (1) SNP1 (NC_030835.1:g.15498308) locus: As shown in Table 2, different genotypes of the SNP1 locus on the seventh intron of the goat ADK gene were significantly associated with goat weight and height. Individuals with the AA genotype had significantly better weight (42.36±7.12kg) and height (61.82±4.63cm) than individuals with the AG genotype (34.61±4.29kg, 56.07±2.06cm, P values 0.049 and 0.034, respectively; no significant differences were found in body length, chest circumference, cannon bone circumference, and rump width). This indicates that the A-to-G mutation at this locus is a harmful mutation, and genotype AA is a candidate marker for early selection of growth traits in goats, which can be applied in marker-assisted selection and molecular breeding in goats.
[0092] Table 2. Association analysis between ADK gene SNP1 and growth traits in Anhui white goats.
[0093]
[0094] (2) SNP2 (NC_030835.1:g.15498501) locus: As shown in Table 3, different genotypes of the SNP2 locus on the seventh intron of the goat ADK gene were significantly associated with the height, length and chest circumference of goats. Among them, the height of the GG genotype individuals was 67.06±2.05cm and the chest circumference was 86.78±5.86cm, which were significantly better than those of the CG and CC genotype individuals, with P values of 0.028 and 0.032, respectively. The length of the CC genotype individuals was 75.03±5.51cm, which was significantly higher than that of the CG genotype individuals, which was 73.22±3.59cm, with P value of 0.033. There were no significant differences in weight, cannon bone circumference and rump width. This indicates that the C-to-G mutation at this locus is a beneficial mutation, but the heterozygous genotype corresponds to the worst phenotype. Therefore, the GG genotype should be selected as a candidate marker for early selection of growth traits in goats and applied in marker-assisted selection and molecular breeding in goats. Heterozygous individuals at this locus should be eliminated as soon as possible.
[0095] Table 3 Association analysis of ADK gene SNP2 and growth traits in Anhui white goats.
[0096]
[0097] (3) SNP3 (NC_030835.1:g.15498816) locus: As shown in Table 4, different genotypes of the SNP3 locus on the seventh intron of the goat ADK gene are significantly associated with body length and chest circumference in goats. Among them, individuals with the TT and GG genotypes have significantly better body length than individuals with the TG genotype, while individuals with the GG genotype have significantly better chest circumference than individuals with the TT and TG genotypes. This indicates that the T-to-G mutation at this locus is a beneficial mutation. However, the heterozygous genotype corresponds to the worst phenotype. Therefore, the GG genotype should be selected as a candidate marker for early selection of growth traits in goats and applied in marker-assisted selection and molecular breeding in goats. Heterozygous individuals at this locus should be eliminated as soon as possible.
[0098] Table 4. Association analysis of ADK gene SNP3 and growth traits in Anhui white goats.
[0099]
[0100] Example 6:
[0101] A method for molecular marker-assisted selection breeding of Anhui White Goats is disclosed. This invention employs the method described herein for detecting SNPs in the ADK gene of Anhui White Goats. The genotypes of three SNP loci at the seventh intron of the ADK gene in candidate breeding goats are detected. Individuals with the AA genotype at locus NC_030835.1:g.15498308, the GG genotype at locus NC_030835.1:g.15498501, and the GG genotype at locus NC_030835.1:g.15498816 are selected to form a core breeding population for the genetic improvement of growth traits in Anhui White Goats.
[0102] The above is an exemplary description of the invention. Obviously, the specific implementation of the invention is not limited to the above-described manner. Any non-substantial improvement made using the inventive concept and technical solution of the invention, or the direct application of the inventive concept and technical solution to other situations without modification, is within the protection scope of the invention.
Claims
1. A method for detecting ADK gene SNPs in Anhui white goats, characterized in that, Using the whole genome DNA of the Anhui white goat as a template, the seventh intron fragment of the ADK gene was obtained by PCR amplification. After the band was confirmed to be correct by agarose gel electrophoresis, Sanger sequencing was performed to identify three SNP sites, namely NC_030835.1:g.15498308, NC_030835.1:g.15498501, and NC_030835.1:g.15498816. The primers used for the PCR amplification are: Upstream primer: 5'-TGATGACCTGCATTAAGCACCA-3'; Downstream primer: 5'-AGCCATAATCCGATCTGAGGTAA-3'.
2. The method for detecting ADK gene SNPs in Anhui white goats according to claim 1, characterized in that, The PCR amplification reaction procedure is as follows: pre-denaturation at 93–95 ℃ for 3–5 min; denaturation at 95–98 ℃ for 10–30 s, annealing at 55–60 ℃ for 30–50 s, extension at 65–72 ℃ for 1–2 min, for 30–35 cycles; extension at 65–72 ℃ for 3–5 min, and storage at 4 ℃.
3. The method for detecting ADK gene SNPs in Anhui white goats according to claim 1, characterized in that, The PCR amplification reaction system includes: 12.5 μL of 2×PCR MasterMix, 25–50 ng of whole genome DNA of Anhui white goats to be tested, and 2.5–5 pmol each of upstream and downstream primers for amplification, with ultrapure water added to bring the total volume to 25 μL.
4. The method for detecting ADK gene SNPs in Anhui white goats according to claim 1, characterized in that, The agarose gel electrophoresis was performed using an agarose gel with a mass fraction of 1.5%–2.0%, and the PCR amplification product fragment size was 767 bp.
5. The method for detecting ADK gene SNPs in Anhui white goats according to claim 1, characterized in that, The genotype determination rules for Sanger sequencing are as follows: The NC_030835.1:g.15498308 locus has the genotype AA when it is a single peak and the genotype AG when it is a multi-peaked locus. At the NC_030835.1:g.15498501 locus, the genotype is CC or GG when there is a single peak, and CG when there are multiple peaks. The NC_030835.1:g.15498816 locus has a genotype of TT or GG when it is a single peak and a genotype of TG when it is a multi-peak.
6. A method for molecular marker-assisted selection breeding of Anhui white goats, characterized in that, The method for detecting SNPs in the ADK gene of Anhui white goats according to any one of claims 1 to 5 is used to detect the genotypes of three SNP sites in the intron 7 of the ADK gene of Anhui white goats, and the three SNP sites are used as molecular markers to screen individuals with excellent growth traits.
7. The method for molecular marker-assisted selection breeding of Anhui white goats according to claim 6, characterized in that, The growth traits include body weight, height, length, chest circumference, cannon circumference, and rump width.
8. The method for molecular marker-assisted selection breeding of Anhui white goats according to claim 6, characterized in that, Anhui white goats with the AA genotype at NC_030835.1:g.15498308, the GG genotype at NC_030835.1:g.15498501, and the GG genotype at NC_030835.1:g.15498816 were selected as the core breeding group.