Chirgwin J M, Przybyla A E, MacDonald R J, Rutter W J
Biochemistry. 1979 Nov 27;18(24):5294-9. doi: 10.1021/bi00591a005.
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.
通过在含有强效蛋白质变性剂硫氰酸胍的4M溶液加0.1M 2-巯基乙醇中进行高效匀浆,以断裂蛋白质二硫键,已从富含核糖核酸酶的组织(如大鼠胰腺)中制备出完整的核糖核酸(RNA)。通过乙醇沉淀或通过氯化铯沉降,分离出不含蛋白质的RNA。通过这些方法获得的大鼠胰腺RNA已用作纯化α-淀粉酶信使核糖核酸的来源。