Glissin V, Crkvenjakov R, and Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13: 2633-2637

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摘要：

A quick and simple procedure based upon CsCl ultracentrifugation for the isolation of RNA from disrupted cells or subcellular fractions is described. rRNA is obtained intact and can be recovered in yields close to 100%. It is shown that the translation of rabbit globin mRNA is not impaired by exposure to this treatment. The yield of poly(A) containing globin mRNA extracted by this method and partially enriched on a poly(U)-Sepharose column was assayed in wheat germ S-30 extracts along with that extracted with phenol undergoing identical treatment. RNA from the CsCl extraction yields RNA six times more active in globin synthesis. It is observed that specific aggregation takes place during CsCl centrifugation at high gravitational fields diminishing the amount of the 16S or 18S rRNA component but not 26S or 28S component. The aggregation can be avoided by slow-speed centrifugation or reversed by heating at 60° for 30 sec. The CsCl centrifugation appears to be superior to phenol extraction in terms of simplicity, processing time, yield, intactness, and biological activity of resulting RNA. The suitability of the method for small amounts of starting material and one step separation of macromolecules from each other seem to recommend the CsCl centrifugation as a general method of isolating RNA over other methods currently in use.

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DOI：

10.1021/bi00709a025

被引量：

1961

年份：

1974