一种简单经济的提取水稻未成熟种子RNA的方法

›› 2007, Vol. 9 ›› Issue (4): 111-115.

一种简单经济的提取水稻未成熟种子RNA的方法

宋贵生翟红利魏刚朱祯

中国科学院遗传与发育生物学研究所,北京100101

收稿日期:2007-05-15 修回日期:2007-07-20 出版日期:2007-08-15 发布日期:2009-10-16

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

SONG Gui-sheng,ZHAI Hong-li, WEI Gang,ZHU Zhen

Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China

Received:2007-05-15 Revised:2007-07-20 Online:2007-08-15 Published:2009-10-16
About author:Song Gui-sheng|Ph．D．candidate in Molecular Genetics．Corresponding author：Zu Zhen|professor in plant molecular genetics．Tel：86—10-64873490：E．mail：zzhu@ genetics．ac．cn
Supported by:
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 Transgenic Plants （J99-B-004,J99-A-020） and Rockefeller Foundation （RF97001#581）.

摘要/Abstract

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520 μg到800 μg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.

关键词: 未成熟种子多糖 RNA提取

Abstract:

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.

Key words: immature seeds;polysaccharides;RNA extraction

中图分类号:

Q522

宋贵生翟红利魏刚朱祯. 一种简单经济的提取水稻未成熟种子RNA的方法[J]. , 2007, 9(4): 111-115.

SONG Gui-sheng,ZHAI Hong-li, WEI Gang,ZHU Zhen . A Simple and Inexpensive Total RNA Extraction Method from the Rice Immature Seeds Rich in Polysaccharides[J]. , 2007, 9(4): 111-115.

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