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改进的蛋白质和核酸同时分离方法。

Improved method for simultaneous isolation of proteins and nucleic acids.

机构信息

Institute of Virology, University of Leipzig, Leipzig, Germany.

出版信息

Anal Biochem. 2011 Apr 1;411(1):164-6. doi: 10.1016/j.ab.2010.11.020. Epub 2010 Nov 19.

Abstract

Guanidinium thiocyanate-phenol-chloroform extraction (GTPC extraction) is widely used in molecular biology for isolating DNA, RNA, and proteins. Protein isolation by commercially available GTPC solutions is time consuming and the resulting pellets are only incompletely soluble. In this study ethanol-bromochloropropane-water was used for precipitation of proteins from the phenol-ethanol phase after GTPC extraction of RNA and DNA. The precipitated proteins can be readily dissolved in 4% SDS for subsequent analysis. This technique allows a fast (30min) and efficient (with a protein recovery of up to 95%) extraction of proteins for the study of transcriptional and posttranscriptional events from the same sample.

摘要

胍盐-酚-氯仿抽提(GTPC 抽提)广泛应用于分子生物学中,用于分离 DNA、RNA 和蛋白质。使用市售的 GTPC 溶液进行蛋白质分离非常耗时,而且得到的沉淀只不完全溶解。在这项研究中,在 GTPC 提取 RNA 和 DNA 之后,使用乙醇-溴氯丙烷-水从酚-乙醇相中沉淀蛋白质。沉淀的蛋白质可以很容易地溶解在 4%SDS 中,以便进行后续分析。该技术允许从同一样品中快速(30min)且高效(蛋白质回收率高达 95%)提取蛋白质,用于研究转录和转录后事件。

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