Nucleic Acids Research, 1979, Vol. 7, No. 6 1513-1523
© 1979
Articles |
A rapid alkaline extraction procedure for screening recombinant plasmid DNA
Laboratoire de Génétique Moléculaire, Institut de Recherche en Biologie Moleculaire, F-75221 Paris Cedex 05, France
+Send correspondence to, H. C. Birnboim,Atomic Energy of Canada Limited,Chalk River, Ontario K0J lJ0 (Canda).
Received August 3, 1979. A procedure for extracting plasmid DNA from bacterial cells 1s described. The method 1s simple enough to permit the analysis by gel electrophoresis of 100 or more clones per day yet yields plasmid DNA which is pure enough to be digestible by restriction enzymes. The principle of the method is selective alkaline denaturation of high molecular weight chromosomal DNA while covalently closed circular DNA remains double-stranded. Adequate pH control is accomplished without using a pH meter. Upon neutralization, chromosomal DNA renatures to form an insoluble clot, leaving plasmid DNA in the supernatant. Large and small plasmid DNAs have been extracted by this method.
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