生物技术通报2017，Vol.33Issue(11)：60-66,7.DOI:10.13560/j.cnki.biotech.bull.1985.2017-0319
一种大量提取红豆杉中总RNA的方法研究
A Method for Extracting Total RNA of Taxus chinensis at Large-scale
摘要
Abstract
This study was directed to develop the optimal method for the extraction and purification of total RNA in Taxus chinensis R.First,we investigated the effects of extraction reagent,time,and method on the yield of total RNA from T.chinensis R.using Trizol and CATB extraction methods in respect of RNA concentration and the ratios of OD260/OD280 and OD260/OD230.Furthermore,we compared the effects of PVP,β-mercaptoethanol,high salt precipitation,low concentration ethanol precipitation and gel column chromatography on the purification of total RNA.The optimal extraction method was determined as follows:treatment of taxus leaves with Trizol by the action volume of 3:40 in 5 minutes using chloroform and water (1:5) as solvent.It's also found that column chromatography significantly improved the purity of total RNA of taxus and obtained small molecular RNA with rich miRNA.Finally,the high-quality RNA in concentration of 1000μg/mL with 2.00-2.20 of OD260/OD280 and 1.7 of OD260/OD230was acquired.Compared with previous methods,this method provides taxus RNA in large-scale with high quality and purity as well as with low cost,which guarantees sufficient materials.关键词
红豆杉/总RNA提取/Trizol法/CATB法/凝胶柱层析法Key words
Taxus chinensis R./total RNA extraction/Trizol method/CATB method/gel column chromatography引用本文复制引用
陈小凤,曾奇峰,邓旭,曾光尧,周应军..一种大量提取红豆杉中总RNA的方法研究[J].生物技术通报,2017,33(11):60-66,7.基金项目
国家自然科学基金项目(81573314) （81573314）
