Chomczynski P
College of Medicine, University of Cincinnati.
Biotechniques. 1993 Sep;15(3):532-4, 536-7.
This report describes a new method for simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. The method is based on the use of a reagent containing phenol and guanidine thiocyanate. A biological sample is homogenized in the reagent and the simultaneous isolation of RNA, DNA and proteins is accomplished in a single step by a liquid-phase separation. The isolation of RNA can be completed in about 1 h, and DNA and proteins in about 3 h. The simultaneously isolated RNA, DNA and proteins are ready for Northern, Southern and Western blotting. The complete recovery of DNA from samples used for the RNA and protein isolation makes it possible to normalize the results of gene expression studies based on DNA content instead of on the more variable total RNA, protein content or tissue weight.
本报告描述了一种从细胞和组织样本中同时分离RNA、DNA和蛋白质的新方法。该方法基于使用一种含有苯酚和硫氰酸胍的试剂。将生物样本在该试剂中匀浆，通过液相分离在一步中同时完成RNA、DNA和蛋白质的分离。RNA的分离可在约1小时内完成，DNA和蛋白质的分离可在约3小时内完成。同时分离得到的RNA、DNA和蛋白质可直接用于Northern、Southern和Western印迹分析。从用于RNA和蛋白质分离的样本中完全回收DNA，使得基于DNA含量而非更易变的总RNA、蛋白质含量或组织重量来标准化基因表达研究结果成为可能。
