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

摘要:

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.

This study is aimed to obtain high-quality total RNA from Dioscorea opposita Thunb.. Total RNA from nodal segments of D. opp. cv. Tiegun was extracted by the trizol reagent, the RNAiso for Polysaecharide-rich Plant Tissue Kit method, a modified CTAB method, and the TaKaRa MiniBEST Plant RNA Extraction Kit method. The quality of isolated to- tal 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 Extrac tion 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.

作者:

李书杰 郭晓博 王运英 赵喜亭 李俊华 李明军

机构地区:

betway官方app 生命科学学院 betway官方app 河南省绿色药材生物技术工程实验室 betway官方app 河南省高校道地中药材保育及利用工程技术研究中心

出处:

《betway官方app 学报：自然科学版》 CAS 北大核心 2015年第3期129-132,共4页

基金:

国家自然科学基金(81274019) 河南省创新型科技人才队伍建设工程(C20130037) 新乡市科技创新平台建设项目(CP1401) betway官方app 青年科学基金项目(2013QK11)

关键词:

山药 茎段 总RNA提取 多酚类物质 多糖

Dioscorea opposita Thunb. stem segment total RNA isolation polyphenol-rich plants polysaccharide

分类号:

Q946 [生物学—植物学]

山药总RNA不同提取方法的比较研究.pdf