雷公藤叶片基因组DNA不同提取方法比较

目的]比较雷公藤叶片中基因组DNA的提取方法。方法]采用常规CTAB法、改良CTAB法、常规SDS法、高盐低pH值法和一步法分别提取雷公藤叶片基因组DNA,利用琼脂糖凝胶电泳、紫外分光光度法测定样品OD260与OD280的比值以及RAPD扩增结果,判断所得DNA样品的纯度,并根据OD260值计算不同提取方法的DNA得率。结果]改良CTAB法提取效果最佳,所提DNA的平均得率为219.3μg/g,OD260/OD280比值在1.869～1.903,用于RAPD-PCR均有较好的扩增结果。结论]改良CTAB法是雷公藤叶片基因组DNA提取的最佳方法。

Comparasion on Several Extraction Methods of Genomic DNA from Tripterygium wilfordii Hook.f.Leaves

Objective] Comparison of the extraction methods of genomic DNA from the leaves of Tripterygium wilfordii Hook.f..Method] With leaves of Tripterygium wilfordii Hook.f.as tested material,the genomic DNA was extracted using five different methods:CTAB method,modified CTAB method,SDS method,high salt and low pH method and ROSE method.The purity of the DNA was examined by agarose gel electrophoresis,OD260/OD280 and RAPD amplification,and the yield of the DNA was estimated by OD260.Result] Modified CTAB method was the optimum one for DNA extraction.The ratio of OD260 and OD280 of the genomic DNA extracted by modified CTAB method was 1.869-1.903,and showed good performance in RAPD analysis.Conclusion]The modified CTAB method is an optimum method for the extraction of genomic DNA from Tripterygium wilfordii Hook.f..