Method for identifying meat production traits of sheep and special primer thereof
A technology for sheep and traits, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., to achieve high application value, increase meat production, and high accuracy
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Embodiment 1
[0020] Determination of embodiment 1 SNP site
[0021] 1. Selection of experimental materials
[0022] Collect 40 muscle tissues of Hu sheep and 39 muscle tissues of Suffolk sheep (Xinao Animal Husbandry Co., Ltd., Manas County, Changji, Xinjiang), and extract genomic DNA respectively, and adjust the DNA concentration of each sample to 100ng / μL, a total of 40 DNA samples of Hu sheep and 39 DNA samples of Suffolk sheep were obtained, which were stored at -80°C for future use.
[0023] 2. Design and synthesis of primers
[0024] Since the mature sequence of the sheep miR-1 gene has not been published, the mature sequence of the bovine miR-1 gene (MIMAT0009214, ID: bta-miR-1) was retrieved from the miRBase database (http: / / www.mirbase.org / ) , and compared with the NCBI sheep genome database (http: / / www.ncbi.nlm.nih.gov), and intercepted the 200bp sequences on the left and right flanks, and designed a pair of primers (primer combination) to clone sheep miR-1 Precursor gene, the...
Embodiment example 2m
[0035] Implementation case 2 Establishment of asymmetric PCR-SSCP detection method for miR-1 precursor gene
[0036] 1. PCR amplification
[0037] The genomic DNA of the sheep to be tested obtained in step 1 was used as a template, and U and D were used as primers to carry out PCR amplification to obtain PCR amplification products. PCR amplification system: 1 μL of 100ng genomic DNA, 10 μL of 2×Taq PCR MasterMix, 0.5 μL of forward primer (concentration of 10 μmol / L), 0.5 μL of reverse primer (concentration of 10 μmol / L), add ddH2O to 20 μL. PCR amplification program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s; annealing at 60°C for 30 s; extension at 72°C for 50 s, a total of 30 cycles; final extension at 72°C for 10 min, detected by 1% agarose gel electrophoresis.
[0038] 2. Asymmetric PCR amplification conditions
[0039] The second PCR method is used, that is, the first PCR amplification is performed with primers of equal concentration to amplify t...
Embodiment example 3
[0045] Implementation Case 3 Correlation Analysis of Sheep miR-1 Precursor Gene g.53971777A>G Polymorphism and Meat Production-related Traits of Sheep
[0046] In order to determine whether the g.53971777A>G polymorphism site is related to the meat production of sheep, 40 Suffolk sheep and 39 Hu sheep were used as experimental materials, their genotypes were determined, and the relationship between the gene mutation site and the meat production Quantitative correlation analysis. The genotype and allele frequency of the g.53971777A>G polymorphism of the sheep miR-1 precursor gene in the population were calculated by SPSS13.0 software, and the chi-square test was used to analyze whether the genotype in each population was in the Hardy- Weinberg balance. The results are shown in Table 1.
[0047] The distribution and Hardy-Weinberg equilibrium of the gene frequency and genotype frequency of the SNP site of miR-1 gene in various sheep breeds
[0048]
[0049] Note: The numbe...
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