适于ISSR-PCR扩增的松毛虫基因组DNA的提取

DNA extraction of pine caterpillars for ISSR-PCR reaction

为了方便稳定地获得松毛虫基因组DNA,寻找适合于松毛虫ISSR-PCR扩增的DNA的提取方法,分别用SDS-蛋白酶K法和CTAB法对松毛虫雌成虫进行基因组DNA的提取。结果显示:用SDS-蛋白酶K法提取的基因组DNA带型整齐,无降解,DNA得率和纯度都较高,操作简单无需纯化,用于ISSR扩增时其稳定性和重复性都很好;而用CTAB法提取出的DNA浓度低,杂质较多,琼脂糖凝胶电泳甚至检测不出,此方法提取的DNA作为ISSR反应模板时需要进行纯化。

Genomic DNA were extracted from the pine caterpillars （Dendrolimus） ： D. tabulaeformis, D. spectabilis and D. superans by SDS-Proteinase K method and CTAB method. Each DNA sample was tested by 0.8 % agarose gel following electrophoresis. The results showed that the two methods were useful in genomic DNA extraction of pine caterpillars to some degree. But comparatively, the quality of DNA samples isolated by SDS - Proteinase K was obviously better and more suitable for ISSR - PCR than by CTAB method. DNA extracted by CTAB method must be purified when being used for ISSR - PCR reaction because of more impurity and low concentration.