Hemifusus tuba microsatellite loci, and primers and application of hemifusus tuba microsatellite loci
A technology of microsatellite loci and snails, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve problems such as the destruction of natural resources of snails in sea areas
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[0019] 1. Screening of microsatellite loci
[0020] 1. Genomic DNA extraction
[0021] 1) Mince the snail gastropod muscle frozen at -80°C with a scalpel, quickly weigh about 100 mg and put it into a 1.5 ml sterilized centrifuge tube;
[0022] 2) Add 500 μL of CTAB lysis buffer and 10 μL of proteinase K (20 mg / ml) to each centrifuge tube, digest in a 56-degree water bath overnight, and mix well by turning over every few hours;
[0023] 3) After the sample is completely digested, add an equal amount of CIA (chloroform: isoamyl alcohol = 24:1), rotate for 20 minutes, centrifuge at 10,000 rpm for 10 minutes at room temperature, pipette the supernatant into another clean centrifuge tube (the middle layer cannot be sucked out) Repeat the CIA extraction once; 4) Add PCI (phenol: chloroform: isoamyl alcohol = 25:24:1), the same amount as the supernatant taken, rotate for 10 min, centrifuge at 10,000 rpm for 10 min at room temperature, take the supernatant and clean and sterilize it ...
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