Emmett M, Petrack B
Research Department, CIBA-GEIGY Corporation, Summit, New Jersey 07901.
Anal Biochem. 1988 Nov 1;174(2):658-61. doi: 10.1016/0003-2697(88)90069-3.
A rapid procedure for the isolation of total RNA from small amounts of mammalian tissue (35 to 150 mg) is described. Tissues were homogenized in the presence of RNase inhibitors but in the absence of strong detergents. Contaminants were removed by phenol/chloroform extraction and Sephadex column chromatography. Total RNAs were precipitated with ethanol and sodium acetate. The RNAs isolated were intact and suitable for mRNA quantitation via Northern blot or slot-blot analyses. This procedure isolates total RNAs in high yield and purity, without CsCl ultracentrifugation, and is especially useful when mRNAs must be quantitated from many samples.
本文描述了一种从少量哺乳动物组织（35至150毫克）中快速分离总RNA的方法。组织在存在核糖核酸酶抑制剂但不存在强去污剂的情况下匀浆。通过苯酚/氯仿萃取和葡聚糖凝胶柱色谱法去除污染物。总RNA用乙醇和醋酸钠沉淀。分离得到的RNA完整，适用于通过Northern印迹或狭缝印迹分析进行mRNA定量。该方法无需氯化铯超速离心即可高产率、高纯度地分离总RNA，在必须从多个样品中定量mRNA时特别有用。
