Optimization and comparison of extraction methods of mitochondrial DNA of Oncomelania hupensis
10.16250/j.32.1374.2016228
- VernacularTitle:钉螺线粒体基因组DNA提取方法的优化及比较
- Author:
Kang WANG
;
Shujun XU
;
Yixiao WANG
;
Wenjie CHEN
;
Yinan WANG
;
Fenfang YU
;
Shaosheng WANG
;
Entao SUN
;
Shulin ZHOU
- Keywords:
Oncomelania hupensis;
Mitochondrial genome DNA(mtDNA);
Improved high-concentration-salt precipitation method
- From:
Chinese Journal of Schistosomiasis Control
2017;29(2):159-162
- CountryChina
- Language:Chinese
-
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 pre-cipitation method. The mtDNA samples were identified by using spectrophotometry,agarose gel electrophoresis and the amplifi-cation products of COX1. The nuclear DNA contamination was tested by the amplification products of ITS. Results The concen-tration and yield of the improved method was significantly higher than those of the traditional method(F=3032.65,10185.00, both P<0.01). The mtDNA samples extracted were essentially free of nuclear DNA and protein,meeting PCR,sequence analy-sis and other molecular biology research requirements. Conclusions The improved high-concentration-salt precipitation meth-od for isolating mtDNA is simple,and it has high yield and low cost. The extracted mtDNA can meet relevant analysis require-ments.
