Primers and screening method of crassostrea hongkongensis triplet-base repetition microsatellite markers
A technology of microsatellite markers and Hong Kong oysters, applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve problems such as long breeding cycle, increase application prospects and scope, and death, and achieve PCR expansion Increase product stability, good polymorphism effect
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[0028] 1. Construction of Hong Kong oyster three-base repeat microsatellite DNA enrichment library
[0029] 1.1 Extraction and restriction digestion of genomic DNA
[0030] Take 30 mg of Hong Kong oyster adductor muscle tissue fixed with 70% alcohol, soak it in deionized water 3 times, and change the water after 15 minutes each time to completely remove the alcohol remaining in the muscle. Use clean filter paper to absorb water and put it into a 1.5mL centrifuge tube, then add 400μl lysate and 10μl proteinase K (10mg / ml), and digest in a water bath at 55°C for 3 to 5 hours. Add an equal volume of saturated phenol (200μL), chloroform / isoamyl alcohol (24:1) (200μL) mixture and extract three times. The DNA was precipitated with 1 mL of absolute ethanol, washed with 70% alcohol, dried at room temperature and dissolved in 100 μL of ultrapure water to obtain the genomic DNA of Hong Kong oyster. UV spectrophotometer was used to detect DNA concentration and purity, and 1% agarose gel ele...
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