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一种提取高质量红树植物总RNA的有效方法
引用本文:陈宏林,王玺,何振艳.一种提取高质量红树植物总RNA的有效方法[J].中国生物工程杂志,2010,30(6):84-88.
作者姓名:陈宏林  王玺  何振艳
作者单位:1.沈阳农业大学农学院 沈阳 110161 2.中国科学院植物研究所光合作用与环境分子生理学重点实验室 北京 100093
基金项目:国家"863"计划资助项目 
摘    要:为了从富含单宁和多糖的红树植物中获得高质量的RNA,在总结及改良前人工作的基础上,优化了提取缓冲液的条件,采用碱性条件、PVP及β-巯基乙醇抑制单宁的氧化;利用RNA与单宁、多糖在不同浓度的2-丁氧乙醇中溶解度不同的特性来去除单宁和多糖;同时利用LiCl来选择性沉淀RNA,获纯度较高的RNA可以直接用于cDNA合成、cDNA文库的构建以及RACE等分子生物学操作。

关 键 词:秋茄  红海榄  榄李  红树植物  单宁  多糖  RNA提取  
收稿时间:2010-01-12
修稿时间:2010-04-10

An Effective Method of High Quality Total RNA Isolation of the Mangrove Plants
CHEN Hong-lin,WANG Xi,HE Zhen-yan.An Effective Method of High Quality Total RNA Isolation of the Mangrove Plants[J].China Biotechnology,2010,30(6):84-88.
Authors:CHEN Hong-lin  WANG Xi  HE Zhen-yan
Institution:1.College of Agronomy, Shenyang Agricultural University, Shenyang 110161, China 2.Key Laboratory of Plant Photosynthesis and Molecular Environmental Physiology, Institute of Botany,Chinese Academy of Sciences, Beijing 100093, China
Abstract:In order to get high quality RNA from mangrove plants rich in tannins and polysaccharides,based on summing up and improved our predecessors, the conditions of extraction buffer was optimized to inhibit the oxidation of tannins by the conditions of alkaline conditions, PVP and β-mercaptoethanol. Most of the polysaccharjdes and tannins extracted with nucleic acids are removed by taking advantage of differences in solubility of these compounds in the solvent 2-butoxvethano. Moreover, LiCl was used to selectively precipitate RNA and obtained higher purity of the RNA which can used for operation of molecular biology such as cDNA synthesis, cDNA library construction and RACE technique.
Keywords:Kadelia candel  Rhizophora stylosa  Lumnitzera racemosa  Mangrove  tannins    Polysaccharides  RNA isolation
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