ABSTRACT
OBJECTIVE: To optimize the extraction methods of mitochondrial genome DNA (mtDNA) of Oncomelania hupen- sis. METHODS: The pyrolysis, protein K variable-temperature digestion and high-concentration potassium acetate purification were applied to optimize the high-concentration-salt precipitation method, and then the optimized method was compared with two common extraction methods, the sucrose density gradient centrifugation method and traditional high-concentration-salt precipitation method. The mtDNA samples were identified by using spectrophotometry, agarose gel electrophoresis and the amplification products of COX1. The nuclear DNA contamination was tested by the amplification products of ITS. RESULTS: The concentration and yield of the improved method was significantly higher than those of the traditional method (F = 3 032.65, 10 185.00, both P < 0.01). The mtDNA samples extracted were essentially free of nuclear DNA and protein, meeting PCR, sequence analysis and other molecular biology research requirements. CONCLUSIONS: The improved high-concentration-salt precipitation method for isolating mtDNA is simple, and it has high yield and low cost. The extracted mtDNA can meet relevant analysis requirements.
