一种简单经济的提取水稻未成熟种子RNA的方法(英文)  

A Simple and Inexpensive Total RNA Extraction Method from the Rice Immature Seeds Rich in Polysaccharides

作　　者：宋贵生[1] 翟红利[1] 魏刚[1] 朱祯[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《中国农业科技导报》2007年第4期111-115,共5页Journal of Agricultural Science and Technology

基　　金：National High Science and Technology Program(863)(2001AA212041,2001AA222251);Project of National Natural Science Foundation of China(39989001);National Special Program for Research and Industrialization of TransgenicPlants(J99-B-004,J99-A-020);Rockefeller Foundation(RF97001#581).

摘　　要：多糖污染是从田间水稻叶片和未成熟种子等高含糖组织中提取高质量RNA的主要限制因素。我们介绍一种简单经济的RNA提取方法,它适合从水稻叶片和未成熟种子这些富含多糖的组织中提取总RNA。这种方法可以克服产率低和质量低的问题。RNA提取后经过电泳和分光光度计分析结果表明,每克水稻组织总RNA产率在520μg到800μg之间,OD260/230比值大于2.0,OD260/280比值大于1.9。提取的RNA可以用于体外反转录cDNA、mRNA分离和基因表达系列分析(serial analysis of gene expression,SAGE)库的构建。Polysaccharides contamination is the major limitative factor for high-quality RNA extraction from the high level polysaccharides tissues in field-grown rice leaves and immature seeds. Here we introduce a simple and inexpensive method for total RNA extraction. It is suitable for extracting total RNA from rice leaves and immature seeds rich in polysaccharides . This method overcame the shortcoming of low yield and low quality . After extraction, the RNA was analyzed by spectrophotometric readings and electrophoresis in denaturing agarose gels. The result indicates that total RNA yield is ranging from 520 μg to 800 μg per gram of tissue with an averaged purity measured as OD260/230 of more than 2.00 and OD260/280 more than 1.900. The extracted RNA can be used in exclusive amplification to cDNA template RT-PCR, mRNA isolation and SAGE （serial analysis of gene expression） library construction.

关 键 词：未成熟种子 多糖 RNA提取 

分 类 号：Q522[生物学—生物化学]