鱼腥草叶片总RNA提取方法的比较与分析

摘要

Objective To establish an effective method for the isolation of the high-quality RNA from Folium Houttuyniae. Methods The total RNA from Folium Houttuyniae was isolated respectively by four methods including RNAiso for Polysaccharide-rich Plant Tissue, cetyltrimethyl ammonium bromide ( CTAB) method, sodium dodecyl sulphate (SDS) method 1 and SDS method 0 . Six reagent combinations were selected to extract the supernatant of homogenale. The purity, productivity and integrity of RNA were analyzed by macroscopic appearance observation, UV absorbance detection and agarose gel electrophorography. And then the effectiveness of various isolation methods was compared. Results The purity and integrity of total RNA were satisfactory by using the methods of RNAiso for Polysaccharide-rich Plant Tissue, CTAB and SDS II together with Phenol-chloroform-isoamyl alcohol (PCI) or PCI combined with NaCl for the extraction. The maximum productivity was 172. 6 μg/g. Conclusion An effective RNA isolation method for Folium Houttuyniae has been established, which will provide evidencne for the research of transcriptomc of Herba Houttuyniae.%[目的]建立提取鱼腥草叶片高质量RNA的有效方法.[方法]采用RNAiso for Polysaccharide-rich Plant Tissue法、十六烷基三甲基溴化铵( CTAB)法、十二烷基硫酸钠(SDS)Ⅰ法和SDSⅡ法提取鱼腥草叶片总RNA.在匀浆上清液抽提时分别选用6种试剂组合,通过现象观察,检测紫外吸光度值和琼脂糖凝胶电泳图,考察RNA的纯度、产率和完整性,比较各方法提取效果的差异.[结果]RNAiso for Polysaccharide-rich Plant Tissue法、CTAB法和SDSⅡ法使用苯酚/氯仿/异戊醇或NaCl加苯酚/氯仿/异戊醇抽提后,所得RNA纯度和完整性均好,最大得率为172.6μg/g.[结论]确定了鱼腥草总RNA提取的有效方法,为鱼腥草转录组研究打下基础.