Primers and method for distinguishing Thai marble goby fries from local marble goby fries
A local bamboo shoot shell fish and bamboo shoot shell fish technology, applied in the field of aquaculture, can solve problems such as improper post-processing, and achieve low cost, clear strips, and strong practical effects
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[0023] 1. Extraction of genomic DNA (Cherna chinensis population, Channa spp. population in rivers):
[0024] 1) Take 0.1~0.3 g of the caudal fin of the experimental fish and add 450 μL DNA extraction buffer (10 mmol / L Tris-HCl, 5 mmol / L EDTA, 75 mmol / L NaCl), mix well and then add SDS and proteinase K to The final concentrations were 0.5% and 200 μg / mL, respectively, and then digested overnight in a water bath at 55°C.
[0025] 2) Add an equal volume of saturated phenol, phenol / chloroform / isoamyl alcohol (25:24:1), and chloroform / isoamyl alcohol (24:1) to extract once each. Each extraction was mixed gently for 10 minutes, centrifuged at 12000 rpm for 20 minutes at 4°C, and the supernatant was taken.
[0026] 3) Add 2.5 mol / L NaCl (final concentration: 0.2 mol / L) and pre-cooled 2 times the volume of absolute ethanol to precipitate the obtained supernatant, centrifuge at 12000rpm for 20 minutes to take the precipitate; rinse with 70% ethanol twice Naturally dry at room temper...
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