摘要
[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。
[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..
出处
《安徽农业科学》
CAS
北大核心
2010年第11期5553-5555,共3页
Journal of Anhui Agricultural Sciences
基金
福建省教育厅A类科技项目(JA07170)
关键词
雷公藤
DNA提取
不同提取方法
Tripterygium wilfordii Hook.f..
DNA extraction
Different extraction methods
