Larimichthys polyactis microsatellite DNA molecular marker
A technology of microsatellite markers and DNA molecules, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., can solve the problem of limited quantity
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[0092] 1. Construction of microsatellite molecular marker enrichment library of small yellow croaker:
[0093] 1.1 Genomic DNA extraction:
[0094] Genomic DNA was extracted by the phenol-chloroform method, and the specific steps were as follows: Cut 20 mg of caudal fin into a 1.5 ml centrifuge tube, add 570 μL of STE buffer (10 mmol / L Tris-Cl, pH=8.0; 100 mmol / L EDTA, pH=8.0; 100 mmol / L L NaCl) and cut up the tissue. Add 10μL 20mg / ml proteinase K and 20μL 20% SDS, digest at 55-56℃. Add 600 μL of phenol:chloroform:isoamyl alcohol (25:24:1) to extract twice, centrifuge at 13000r / min for 5min, and take the supernatant. Add an equal volume of -20°C pre-cooled absolute ethanol, mix well, centrifuge at 13000r / min for 5min, discard the supernatant, wash the precipitate twice with 70% ethanol, dry at room temperature, then add 50-100μL TE buffer (1mmol / L EDTA, 10mmol / L Tris-HCl, pH=8.0) to dissolve DNA. The DNA concentration was detected by an ultraviolet spectrophotometer, dilu...
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