香鱼基因组DNA提取方法的优化

为获得高质量的基因组DNA,分别采用传统酚-氯仿法、高盐法、试剂盒法和改进酚氯仿法提取香鱼肌肉基因组DNA。琼脂糖凝胶电泳检测结果表明,改进酚氯仿法提取的基因组DNA电泳条带整齐明亮且无降解。紫外分光度计测定DNA浓度和纯度,结果表明,改进酚氯仿法提取的鱼类基因组DNA浓度约为300μg/mL,A260/A280为1.80-1.86。用改进的酚氯仿法提取的DNA进行AFLP分析,扩增结果稳定,电泳条带清晰。综上所述,改进酚氯仿法能够获得高质量DNA,且可以用于进一步的分子生物学研究。

Extraction of High Quality Genome DNA of Musculature from Ayu ( Plecoglossus altivelis )

In order to obtain high quality genome DNA,the DNA of musculature from ayu(Plecoglossus altivelis) was respectively extracted with methods of traditional phenol-chloroform,high salting out,DNA extraction kit and an improved phenol-chloroform.The genome DNA was appraised by agarose gel electrophoresis,and the result showed that the gel bands by improved phenol-chloroform method were neat,bright,and no degradation.Then,the concentration and purity of DNA were detected by spectrophotometer,and the results showed that the concentration of DNA was approximately 300 μg/mL,and the ratio of A260/A280 was between 1.80-1.86.The genome DNA extracted with improved phenol-chloroform method was used for AFLP analysis,and the result was stable and the electrophoresis banding was clear.So high quality genome DNA can be extracted with improved phenol-chloroform method and used in further study of molecular biology.