辣椒基因组DNA两种提取方法的比较

Comparative Study on Two methods of DNA Extraction from Pepper

采用SDS法和CTAB法提取新鲜辣椒幼叶的基因组DNA,用核酸蛋白仪检测法、琼脂糖凝胶电泳法和PCR扩增法对两种方法提取的DNA样品分别进行检测,核酸蛋白仪检测表明:SDS法得到的DNAOD260/OD280均大于2.0,说明蛋白质去除彻底,但RNA未除干净;CTAB法得到的DNAOD260/OD280均为1.8～2.0,说明蛋白质、多糖、RNA完全去除。琼脂糖凝胶电泳表明:SDS法提取的DNA有两条带,说明样品中仍含有未被除去的多糖及其他一些次生代谢物质,RNA也有残留;CTAB法提取的DNA只有一条清晰整齐的主带,有效地去除了辣椒幼叶中的多糖及其他次生代谢物质,RNA也降解完全。PCR扩增法显示:SDS法和CTAB法提取的DNA样品均可进行PCR扩增,也可用作SRAP研究,但CTAB法提取的样品扩增出的条带比较清晰。结果表明:CTAB法提取的DNA含量高,纯度高,能够满足后续分子生物学研究的需要。

CTAB and SDS method were adopted to extract DNA from fresh pepper leaves. Nucleic acid protein detector method,agarose get electrophoresis method and PCR amplification methods were applied to measure the DNA samples. Nucleic acid protein detector measurement showed OD260/OD280 extracted with SDS method was above 2.0,which means protein has been wiped out completely while RNA has not been wiped out completely yet; OD260/OD280 extracted with CTAB method range from 1.8 to 2.0,which means protein, polysaccharides and RNA have been wiped out completely. Agarose gel electrophoresis measurement showed DNA extracted with SDS method has polysaccharides and RNA which failed to be removed and other secondary metabolites, while polysaccharides,RNA and other secondary metabolites can not be found in DNA extracted with CTAB method. PCR amplification measurement shows both DNA extracted with SDS method and DNA extracted with CTAB method can be amplified and used for SRAP research,but samples extracted with CTAB have clearer amplification band. The results showed DNA extracted with CTAB has high content ,purity,and it can satisfy the needs of research on molecular biology.