Low-temperature-resistant molecular marker A257 of penaeus japonicus and application of molecular marker A257
A technology of molecular markers and low temperature resistance, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of lack of molecular markers, and achieve the effect of accurate and reliable methods, increased quantity, and simple operation
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[0026] 1. Screening of candidate molecular markers related to low temperature resistance
[0027] 1. Sequencing data filtering and comparison
[0028] Genomic DNA of Penaeus japonicus was extracted, 20 individuals in the low temperature tolerance group and 20 intolerant groups. 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 of GA buffer and 20 μL of Proteinase K solution, vortex to mix, and incubate at 56°C until the tissue is completely lysed. Then add 200 μL of GB buffer, mix thoroughly by inversion, place at 70°C for 10 minutes, the solution becomes clear, cut it short and centrifuge to remove water droplets on the inner wall. Add 200 μL of absolute ethanol, invert and mix thoroughly. At this time, flocculent precipitates may appear, centrifuge briefly, and then add the resulti...
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