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从植物组织中分离RNA的改良方法。

Improved method for the isolation of RNA from plant tissues.

作者信息

Logemann J, Schell J, Willmitzer L

出版信息

Anal Biochem. 1987 May 15;163(1):16-20. doi: 10.1016/0003-2697(87)90086-8.

Abstract

A fast and efficient method for the isolation of RNA from plant tissues is described. Tuber tissue is homogenized in a guanidine hydrochloride-containing buffer followed by direct extraction with phenol/chloroform. The RNA is precipitated from the aqueous phase, washed with 3 M sodium acetate and 70% ethanol, and finally dissolved in water. The yield of RNA is up to 500 micrograms/g of tissue and several tests indicate intact and nondegraded RNA. This method can be adapted to a small-scale version by the use of 1.5-ml tubes, allowing rapid isolation of RNA from a larger number of samples. Finally, this method is of particular use for isolating RNA from tissues with a high polysaccharide and nuclease content such as wounded potato tubers.

摘要

本文描述了一种从植物组织中快速高效分离RNA的方法。将块茎组织在含盐酸胍的缓冲液中匀浆,然后用苯酚/氯仿直接提取。RNA从水相中沉淀出来,用3M醋酸钠和70%乙醇洗涤,最后溶解于水中。RNA的产量高达500微克/克组织,多项测试表明RNA完整且未降解。通过使用1.5毫升试管,该方法可适用于小规模操作,从而能从大量样品中快速分离RNA。最后,该方法特别适用于从多糖和核酸酶含量高的组织(如受伤的马铃薯块茎)中分离RNA。

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