Mesh : Chromatography / instrumentation DNA, Bacterial / isolation & purification Electrophoresis, Agar Gel Hydrogen-Ion Concentration Molecular Weight Plasmids RNA, Bacterial / isolation & purification

来  源:   DOI:10.1016/0003-2697(85)90636-0

Abstract:
A procedure for extracting RNA-free plasmid DNA from bacterial cells is described. The method is simple and rapid enough to obtain pure plasmid DNA in 8 to 10 h after plasmid amplification. The protocol uses the alkaline extraction procedure described by Birnboim and Doly (1979, Nucl. Acid Res. 7, 1513-1523). Plasmid DNA is then separated from high-molecular-weight RNA by ammonium acetate precipitation and from low-molecular-weight RNA contaminants by Ultrogel A2 column chromatography. The plasmid DNA obtained by this inexpensive technique is sufficiently pure to be used for restriction endonuclease analysis, 5\'-end labeling, S1 mapping, DNA sequencing, and colony hydridization.
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