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一种从存档的人类血液和肌肉中提取RNA和基因组DNA的简化方案。

A streamlined protocol for extracting RNA and genomic DNA from archived human blood and muscle.

作者信息

Majumdar Gipsy, Vera Santiago, Elam Marshall B, Raghow Rajendra

机构信息

Department of Veterans Affairs Medical Center, Memphis, TN 38104, USA; Department of Pharmacology, College of Medicine, University of Tennessee Health Science Center, Memphis, TN 38163, USA.

Department of Veterans Affairs Medical Center, Memphis, TN 38104, USA.

出版信息

Anal Biochem. 2015 Apr 1;474:25-7. doi: 10.1016/j.ab.2014.12.021. Epub 2015 Jan 9.

Abstract

We combined the TRIzol method of nucleic acid extraction with QIAamp columns to achieve coextraction of RNA and genomic DNA from peripheral blood mononuclear cells (PBMCs) and biopsied skeletal muscle, both stored at -80 °C for many months. Total RNA was recovered from the upper aqueous phase of TRIzol. The interphase and organic phases were precipitated with ethanol, digested with proteinase K, and filtered through QIAamp MinElute columns to recover DNA. The combined protocol yielded excellent quality and quantity of nucleic acids from archived human PBMCs and muscle and may be easily adapted for other tissues.

摘要

我们将TRIzol核酸提取方法与QIAamp柱相结合,以实现从储存于-80°C数月的外周血单个核细胞(PBMC)和活检骨骼肌中同时提取RNA和基因组DNA。从TRIzol的上层水相中回收总RNA。将中间相和有机相用乙醇沉淀,用蛋白酶K消化,然后通过QIAamp MinElute柱过滤以回收DNA。该联合方案从存档的人类PBMC和肌肉中获得了高质量和高产量的核酸,并且可以很容易地适用于其他组织。

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