山药总RNA不同提取方法的比较研究

为提取高质量的山药总RNA,以铁棍山药茎段为材料,采用Trizol法、RNAiso for Polysacchariderich Plant Tissue法、改良CTAB法和试剂盒法提取总RNA,利用琼脂糖凝胶电泳分析所提取RNA的完整性,并用超微量分光光度计分析RNA的纯度.结果显示采用Trizol法难以提RNA,RNAiso for Polysacchariderich Plant Tissue法提取效果不理想,存在DNA和蛋白质污染,而用改良CTAB法和试剂盒法提取到的总RNA纯度高、完整性好,其中,改良CTAB法较试剂盒法价格便宜,但提取过程较烦琐.因此,应根据实验需要选择CTAB法或试剂盒法提取铁棍山药总RNA.本结果对其他品种山药或富含多糖多酚类物质植物的RNA的提取也具有一定借鉴意义.

Comparison of Total RNA Isolated from Dioscorea opposita Thunb.Using Different Methods

This study is aimed to obtain high-quality total RNA fromDioscorea opposita Thunb..Total RNA from nodal segments of D.opp.cv.Tiegun was extracted by the trizol reagent,the RNAiso for Polysaccharide-rich Plant Tissue Kit method,a modified CTAB method,and the TaKaRa MiniBEST Plant RNA Extraction Kit method.The quality of isolated total RNA was analyzed by agarose gel electrophoresis and ultramicro-spectrophotometer,and the efficiency of different methods were compared.The results showed that it was difficult to isolate RNA with Trizol method.RNA isolated with RNAiso for Polysaccharide-rich Plant Tissue Kit was contaminated by DNA and protein.With the TaKaRa MiniBEST Plant RNA Extraction Kit and the modified CTAB method,high quality RNA was obtained.Therefore,the modified CTAB method and the TaKaRa MiniBEST Plant RNA Extraction Kit method are better for total RNA isolation from D.opp.The modified CTAB method is more economic than the TaKaRa MiniBEST Plant RNA Extraction Kit method,however,it is more tedious than the TaKaRa MiniBEST Plant RNA Extraction Kit method.Our study is valuable for total RNA isolation from not only Tiegun yam,but also from other varieties of D.opp.and polyphenol-and polysaccharide-rich plants.