孝顺竹RNA提取方法研究  

A Study on Methods for RNA Isolation from Bamboos

作　　者：杨卫东[1] 王敬文[1] 张金萍[1] 费学谦[1] 

机构地区：[1]中国林业科学研究院亚热带林业研究所,浙江富阳311400

出　　处：《林业科学研究》2005年第6期769-772,共4页Forest Research

基　　金：浙江省自然科学基金资助项目(M303374)

摘　　要：提取纯度高、完整性好、不含DNA或其它杂质的总RNA是进行Northern杂交、cDNA合成、cDNA文库构建、RT-PCR及mRNA差异显示等分子生物学研究的基础.目前提取植物RNA方法有多种,这些方法设计旨在RNA提取过程中清除多糖、多酚等杂质污染.在提取高质量RNA的过程中,植物尤其木本植物通常含有大量的酚类、多糖和其他一些水溶性化合物与RNA共沉淀,影响了RNA功能[1].目前应用较多和较广的有5种方法,商业试剂盒TRIzol,以异硫氰酸胍为代表异硫氰酸酸胍法[1,2]、以阳离子去污剂CTAB代表CTAB法[3,4,5]、热硼酸盐法[6,7],和阴离子去污剂SDS为代表SDS法[8～11]等.结合蛋白质、多糖和多酚去除,以及RNA沉淀形成的这些方法,往往要依据不同植物进行适当改进.Isolation of high quality RNA from bamboos was complicated by high levels of polyphenols and polysaccharides which bind or co-precipitate with RNA. Using TRIzol, modified guanidine isothiocyanate, modified CTAB, modified hot borated and modified SDS method were investigated. Yield and quality were monitored by UV absorbance （A260/A250 and A260/A230） and by native agarose gel, quality and yield of total RNA of five method extracted were compared. Result showed that RNA extraction using modified SDS method resulted in RNA of high quality from five methods investigated, ratio of A260/A280 was over 1.9, and ratio of A260/A230 was over 1.2, this method produced RNA at 560 μg · g^-2 （FW） with no degradation. Modified SDS method is an efficient and effective procedure for the extraction of high quality and yield RNA from bamboos.

关 键 词：孝顺竹 RNA 提取 改良SDS法 

分 类 号：S795.9[农业科学—林木遗传育种]