Abstract:In this study, three genomic DNA extraction methods including phenol-chloroform extraction, boiling lysis and reagent kit methods were compared in terms of extraction efficiency, DNA quantity, as well as the results of White Spot Syndrome Virus (WSSV) detection. The DNA extraction efficiency of three methods, was 101.5, 372.6 and 21.5 ng/μl, respectively. The ratios of OD260/OD280 were in the range of 1.979~2.175 (mean 2.070), 1.699~1.932 (mean 1.796) and 1.784~2.075 (mean 1.951), respectively. The positive rate of WSSV by OIE (Office International Des Epizooties) nested-PCR was 60% (DNA from phenol-chloroform extraction method), 50% (DNA from boiling lysis method) and 70% (DNA from kit method), while it was 100% by quantitative PCR (all DNA from three methods). The DNA copy number of WSSV detected by quantitative PCR ranged from 916.0 to 2.23×106, 63.3 to 1.78×106 and 479.7 to 2.70×106 copies/μl, respectively. The results would provide some reference for standardizing the detection of WSSV.
