Majumdar D, Avissar Y J, Wyche J H
Department of Biology, Brown University, Providence, RI 02912.
Biotechniques. 1991 Jul;11(1):94-101.
A very simple procedure for the simultaneous preparation of genomic DNA and total RNA is described. The procedure is essentially the same for eukaryotes and prokaryotes except for the lysis buffer and can be used for small or large numbers of cells. Mammalian cells are lysed in sodium dodecyl sulfate and bacterial cells are lysed in Triton X-100, both in the presence of EDTA. RNA is obtained in the aqueous phase after phenol (acidic pH):chloroform:isoamyl alcohol extraction. DNA is eluted out of the organic phase (and the interface) into the aqueous phase by increasing the pH with highly basic 1 M Tris solution. The method is extremely rapid for small or large numbers of cells, and several large samples can be processed in one day. The qualities of both nucleic acids are excellent and the yield is high.
本文描述了一种同时制备基因组DNA和总RNA的非常简单的方法。除裂解缓冲液外，该方法对于真核生物和原核生物基本相同，并且可用于少量或大量细胞。哺乳动物细胞在十二烷基硫酸钠中裂解，细菌细胞在Triton X-100中裂解，两者均在EDTA存在下进行。经过苯酚（酸性pH）：氯仿：异戊醇萃取后，RNA存在于水相中。通过用高碱性1M Tris溶液提高pH值，DNA从有机相（和界面）洗脱到水相中。该方法对于少量或大量细胞都极其快速，并且一天内可以处理几个大样本。两种核酸的质量都非常好，产量也很高。
