遗传 ›› 2006, Vol. 28 ›› Issue (5): 583-586.

• 技术与方法 • 上一篇    下一篇

一种简单有效的植物RNA提取方法

张容; 郑彦峰; 吴瑶; 王胜华; 陈放   

  1. 四川大学生命科学学院, 成都 610064
  • 收稿日期:2005-11-08 修回日期:2006-01-16 出版日期:2006-05-10 发布日期:2006-05-10
  • 通讯作者: 王胜华

A Simple and Efficient Method for preparation of Plant RNAs

ZHANG Rong,  ZHENG Yan-Feng,  WU Yao,   

  1. College of Life Science, Sichuan University,Chengdu 610064,China
  • Received:2005-11-08 Revised:2006-01-16 Online:2006-05-10 Published:2006-05-10
  • Contact: WANG Sheng-Hua

摘要: 在提取缓冲液中加入皂土有效地去除了蛋白质并抑制RNAse,建立了一种高效的植物RNA提取方法。以麻疯树幼叶为材料,分别用TRIZOL,异硫氰酸胍法,SDS-KAc法和新创皂土法提取总RNA进行比较和验证,琼脂糖凝胶电泳和紫外光谱分析结果表明,只有皂土法能提出质量高,完整性好的RNA。进一步以皂土法提取的18S rRNA为模板的RT-PCR结果分析表明, 用该法提取的RNA 能用于分子克隆与基因表达分析等后继分子生物学实验。该方法简便,快速,不失为一种经济高效的RNA提取方法。

关键词: 皂土, RNA提取, 麻疯树

Abstract: A new and efficient method for isolation of plant RNAs was developed by adding bentonite into extraction buffer in order to get rid of protein and restrain Rnase. The electrophoretic patterns of nucleic acids and absorbance at 230 nm, 260 nm and 280 nm in a UV-Vis spectrophotometer revealed the extraction with this method can obtain RNAs with good integrity and purity without any apparent DNA contamination from the plant materials rich in with polysaccharide and polyphenol like Jatropha curcas leaves, to which TRIZOL reagent, SDS-KAc solution and Guanidine isothiocyanate solution failed. Furthermore, the result of nuclear gene (18 S rRNA gene) amplified by RT-PCR indicated that the RNAs prepared with this method can meet the needs of most molecular biological experiments including gene cloning and expression analysis.

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