Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease

作者：John M. Chirgwin, Alan Przybyla, Raymond J. MacDonald, William J. Rutter · 发表于：Biochemistry · 年份：1979 · DOI：10.1021/bi00591a005 · 被引用次数：22214 · 研究领域：DNA and Nucleic Acid Chemistry、Analytical Chemistry and Chromatography、Protein Structure and Dynamics

Intact ribonucleic acid (RNA) has been prepared from tissues rich in ribonuclease such as the rat pancreas by efficient homogenization in a 4 M solution of the potent protein denaturant guanidinium thiocyanate plus 0.1 M 2-mercaptoethanol to break protein disulfide bonds. The RNA was isolated free of protein by ethanol precipitation or by sedimentation through cesium chloride. Rat pancreas RNA obtained by these means has been used as a source for the purification of alpha-amylase messenger ribonucleic acid.