Traditional Chinese medicinal material genome DNA extraction method suitable for DNA amplification technology
An extraction method and a technology of traditional Chinese medicinal materials, which are applied in the field of molecular biology, can solve problems such as cumbersome procedures, damage to the health of operators, and complicated operations, and achieve the effects of strong versatility, saving reagents and consumables, and little pollution
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[0033] 1. Materials
[0034] The materials are black snake, money snake, earth dragon, and scorpion; the method is the same as the implementation case 1, and the difference between this implementation case and the implementation case 1 is the use of the black snake specific identification primer WSS-1: 5'-GCGAAAGCTCGACCTAGCAAGGGGACCACA- 3', and WSS-2: 5'-CAGGCTCCTCTAGGTTGTTATGGGGTACCG-3'; PCR reaction system: 25 μL PCR reaction system containing 2.5 μL 10×ExTaq buffer, 2 μL 0.25 mmol / L dNTPs; 0.25 pmol each of upstream primer and downstream primer; 0.8 U Ex taq DNA polymerase, 1 μL of the above DNA template. The reaction was amplified on the 9700 gene amplification instrument of ABI company, and the cycle parameters were pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, extension at 63°C for 30 s, 38 cycles; extension at 72°C for 7 min; storage at 4°C after the reaction was completed. .
[0035] 2. Results
[0036] According to the method used in the presen...
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