一种基于 TRIzol 的快速两步法，用于从拟南芥蒴果和干种子中提取无 DNA 的 RNA。

A rapid TRIzol-based two-step method for DNA-free RNA extraction from Arabidopsis siliques and dry seeds.

机构信息

Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720-3102, USA.

出版信息

Biotechnol J. 2010 Feb;5(2):183-6. doi: 10.1002/biot.200900211.

DOI:10.1002/biot.200900211

PMID:20108272

Abstract

Extraction of high-quality RNA from Arabidopsis seeds has been a challenge. Here we report a two-step TRIzol-based procedure for RNA extraction from Arabidopsis siliques and dry seeds. This procedure employs a modified, high pH (pH 9.5) extraction buffer. High pH plus the addition of either DTT or beta-mercaptoethanol in the extraction buffer effectively inhibits RNase activity during the extraction, and removes most polysaccharides, polyphenols and other insoluble material. TRIzol reagent was subsequently used to purify the RNA. Using this procedure we isolated high-quality DNA-free RNA samples without DNase I treatment from Arabidopsis seeds or siliques in less than 3 h.

摘要

从拟南芥种子中提取高质量的 RNA 一直是一个挑战。在这里，我们报告了一种两步 TRIzol 法从拟南芥蒴果和干种子中提取 RNA。该方法采用改良的高 pH（pH 9.5）提取缓冲液。高 pH 值加上在提取缓冲液中添加 DTT 或β-巯基乙醇可有效地在提取过程中抑制 RNA 酶的活性，并去除大部分多糖、多酚和其他不溶性物质。然后使用 TRIzol 试剂纯化 RNA。使用该程序，我们无需使用 DNA 酶 I 处理，即可在不到 3 小时的时间内从拟南芥种子或蒴果中分离出无 DNA 的高质量 RNA 样品。