A molecular marker g2997 for rapid identification of low temperature tolerant varieties of Penaeus japonicus and its application
A technology of G2997-R and molecular markers, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of lack of molecular markers, and achieve accurate and reliable methods, simple operation, and improved nutrition. The effect of success rate and breeding cycle
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[0029] 1. Screening of candidate molecular markers related to low temperature resistance
[0030] 1. Sequencing data filtering and comparison
[0031] Genomic DNA of Penaeus japonicus was extracted from 20 individuals in each of the low temperature tolerance group and the low temperature intolerant group. Genomic DNA of Penaeus japonicus was extracted by using a centrifugal adsorption column that can specifically bind DNA and a unique buffer system. First, put about 30 mg of tissue samples into a 1.5 mL sterile enzyme centrifuge tube, add 200 μL buffer GA and 20 μL Proteinase K solution, vortex to mix, and incubate at 56°C until the tissue is completely lysed. Then add 200 μL buffer GB, mix well by inverting, place at 70°C for 10 min, the solution becomes clear, and briefly centrifuge to remove water droplets on the inner wall. Then add 200 μL of absolute ethanol, invert and mix thoroughly. At this time, flocculent precipitates may appear, centrifuge briefly, and then add th...
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