Applications of single-stranded nucleotide sequence of INHA gene as animal superovulation molecule marker, and detection method
A single-stranded nucleotide and superovulation technology, which is applied in the application and detection field of animal superovulation molecular markers, can solve the problems of heavy workload, large sample consumption, high cost, etc., and achieves improved efficiency, convenient implementation, and method easy effect
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[0050] 1. Genomic DNA was extracted from the blood of Xiangxi yellow cattle with the Genomic DNA Extraction Kit of Tiangen Biochemical Technology Co., Ltd.
[0051] 2. Partial fragment amplification of INHA gene of Xiangxi yellow cattle:
[0052] Primers:
[0053]
[0054] PCR process:
[0055]
[0056]
[0057] For PCR results, see figure 1 .
[0058] 3. SSCP typing of PCR amplification products of different individuals:
[0059] Mix 3 μl of PCR product with 7 μl of denaturing buffer (98% deionized formamide, 10 mmol / L EDTA (pH8.0), 0.025% xylene cyanol FF, 0.025% bromophenol blue), and use a PCR instrument 95 Denature at ℃ for 10 minutes, and then stand on ice for 5 minutes to avoid renaturation of the denatured product, thereby obtaining denatured single-stranded DNA. The denatured DNA was electrophoresed in a 12% non-denaturing polyacrylamide gel at 140V in a refrigerator at 4°C for 14h. The gel was silver-stained with 0.4% silver nitrate to develop the colo...
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