Method for detecting sow total-number-born character-correlated mutant alleles and reagent kit thereof
A technology of total litter size and detection method, applied in the field of in situ genes, can solve the problems of low selection efficiency, lack of corresponding relationship, and inability to directly know individual genotypes, etc., achieve high application value, improve economic benefits, and low detection costs Effect
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[0029] Embodiment 1, establishment of breeding method
[0030] 1. Establishment of PCR-SSCP detection method and determination of polymorphic sites
[0031] 1. PCR amplification
[0032] Primers were designed according to the pig GenBank Accession Number EU407778 sequence, and the primers were as follows:
[0033] U (upstream primer): 5'-ACTACCCCCTTGTCCCAAAC-3' (sequence 1 of the sequence listing);
[0034] D (downstream primer): 5'-GGACCAA GCCAATCAGAGAA-3' (SEQ ID NO: 2 of the Sequence Listing).
[0035] Large white pigs were selected as experimental materials. Using large white pig genomic DNA as a template, PCR amplification was performed using primers U and D.
[0036] Amplification system: 10×buffer 2.0μL, 25μmol / L Mg2+2.0μL, 10μmol / L dNTPs 0.4μL, 10μmol / L upstream and downstream primers 0.4μL, template DNA 50ng, Taq DNA polymerase 0.3μL, ultra-pure water to 20 μL.
[0037] The PCR reaction conditions were: denaturation at 95°C for 5 min; then denaturation at 95°C f...
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