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从少量真核细胞中同时纯化DNA和RNA。

Simultaneous purification of DNA and RNA from small numbers of eukaryotic cells.

作者信息

Karlinsey J, Stamatoyannopoulos G, Enver T

机构信息

Department of Medicine, University of Washington, Seattle 98195.

出版信息

Anal Biochem. 1989 Aug 1;180(2):303-6. doi: 10.1016/0003-2697(89)90435-1.

Abstract

An extraction procedure for the simultaneous isolation of RNA and DNA from tissue culture cells is described. The procedure is a variation of the guanidium/lithium chloride method for RNA isolation which is rapid, simple, and avoids costly ultracentrifugation equipment. The genomic DNA yielded by this procedure is greater than 50 kb in length and may be readily cleaved by restriction endonucleases. Sufficient DNA for Southern blot analysis, and RNA for Northern blot or nuclease protection analysis, can be obtained from as few as 2 x 10(6) cells, making this method particularly suitable for the genetic screening of large numbers of individual, stably transfected cell clones.

摘要

本文描述了一种从组织培养细胞中同时分离RNA和DNA的提取方法。该方法是胍盐/氯化锂RNA分离方法的一种变体,具有快速、简单的特点,且无需昂贵的超速离心设备。通过该方法获得的基因组DNA长度大于50 kb,并且可以很容易地被限制性内切酶切割。从少至2×10⁶个细胞中就可以获得足够用于Southern印迹分析的DNA,以及用于Northern印迹或核酸酶保护分析的RNA,这使得该方法特别适用于对大量单个稳定转染细胞克隆进行基因筛选。

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