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- W176521568 abstract "The chapter discusses the isolation of soluble RNA from intact E. coli cells. The bacterial paste is deproteinized by phenol treatment. The RNA is precipitated with ethanol to remove low molecular weight material and phenol, sRNA is separated from high molecular weight RNA by extraction with 1 M sodium chloride. Further purification of the sRNA is achieved, either by fractionation with isopropanol or by chromatography on DEAE-cellulose. Isolation operations are carried out in a cold room or at 4° unless specified otherwise. About 60% sRNA is collected. The main impurities are polysaccharides, nucleoside trisphosphates, some DNA, and an RNA fraction, which is inactive as an amino acid acceptor. Then two procedures are used for further purification-fractionation with isopropanol or column chromatography on diethylaminoethyl (DEAE)-cellulose. Both methods achieve about equivalent purification. The commercially available sRNA preparations in some cases are improved considerably by either of the two procedures. The starting material for both procedures is the ethanol precipitate after the step for the removal of the attached amino acids. The chapter also discusses the preparation of 14C- or 3H-aminoacyl-sRNA." @default.
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- W176521568 date "1967-01-01" @default.
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- W176521568 title "[76] Isolation of sRNA from intact Escherichia coli cells" @default.
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- W176521568 doi "https://doi.org/10.1016/s0076-6879(67)12088-0" @default.
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