NGF gene as molecular marker of sheep yeaning traits and application thereof
A molecular marker and sheep technology, which is applied in recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of low selection accuracy, long cycle, and inability to carry out early breeding
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[0030] 1. Blood sample collection
[0031] Peripheral blood samples were collected from sheep with first-born lambing records (breeds Hu sheep and Small-tailed Han sheep), and the blood samples were collected from Jinchang Zhongtian Sheep Industry Co., Ltd., Gansu Province.
[0032] 2. Genomic DNA extraction
[0033] Specific steps are as follows:
[0034] (1) Take 1 mL of collected whole blood and add an equal volume of PBS buffer, and shake gently for 10 minutes;
[0035] (2) After centrifugation at 12000g for 10min at 4°C, discard the supernatant;
[0036] (3) Repeat step (1) and step (2) until the supernatant is transparent and the precipitate turns white;
[0037] (4) Add 700 μL of DNA extraction solution (1M Tris-Cl (pH=8.0) 25mL, 0.5M EDTA (pH=8.0) 100mL, 10% SDS 50mL, 2M NaCl 25mL, set the volume to 500mL, and perform high-pressure Sterilization), suspend the precipitate, and put it in a water bath at 37°C for 1h;
[0038] (5) Add 3 μL proteinase K (20mg / mL) solut...
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