大赖草基因组DNA提取方法比较分析

Comparison of Different Extraction Methods of Genomic DNA in Leymus racemosus(Lam.)Tzvel

为了研究不同提取方法对大赖草[Leymus racemosus(Lam.)Tzvel]基因组DNA提取质量的影响,以大赖草的嫩叶为材料,采用CTAB法、CTAB改良法、SDS法、SDS-CTAB法4种提取方法提取大赖草叶片基因组DNA,对提取的基因组DNA进行琼脂糖凝胶电泳及紫外分光光度计法检测,获得最佳大赖草DNA提取方法 ,并利用ISSR标记验证其质量。结果表明,改良的CTAB法提取的大赖草基因组DNA纯度较高、完整性较好,ISSR扩增条带清晰,多态性高。最终确定CTAB改良法为最佳提取方法,完全适合于进一步的分子生物学研究。

In order to research the effects of different extraction methods on the quality of genomic DNA in Leymus racemo-sus （Lam.） Tzvel and select the optimum method, taking leaves as materials, genomic DNA was extracted by four methods of CTAB,modified CTAB, SDS, and SDS-CTAB, and the quality was tested and verified by agarose gel electrophoresis and ISSR analysis. The results showed that the DNA extracted by the modified CTAB method had a high quality and purity, and was not degraded; in addition,the bands amplified by ISSR-PCR were clear and possessed high polymorphism. In conclusion, the modified CTAB method has the best extraction effect, and could be used for the subsequent molecular biology research.