Spermary-specific protein gene sequence primer and non-electrophoresis method for detecting and identifying cow embryo gender
A specific protein and gene sequence technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as weakness, and achieve the effect of shortening time, improving sensitivity, and not easy to contaminate
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[0015] Put 3-4 drops (each drop 200 μl) of phosphate buffered saline (PBS) in a petri dish, and use MN-151 metal blades to cut the embryos under a stereomicroscope (SZX7-3121, OLYMPUS). About 10% of the inner cell mass of the morula or about 10% of the vegetative ectoderm of the blastocyst are cut for biopsy, and the cut embryos are to be transplanted. Clean the blade with 70% alcohol and sterilized ultrapure water every time an embryo is cut. Six embryos were divided and sampled according to the above method.
[0016] Wash the 6 divided embryo samples with phosphate buffered saline (PBS) for 3 times, respectively draw the divided embryo samples (total volume about 2 μl) with a pipette gun (specification 0.5-10 μl), and transfer them into 0.2ml PCR reaction tubes (6), each PCR reaction tube was pre-put 5 μl alkaline lysate (ALB) (ALB composition: 50mM DTT and 200mM KOH, that is, 50mM dithiothreitol and 200mM potassium hydroxide), 65 ℃ , 10min. Then 5 μl of neutralizing solu...
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