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Nucleic Acids Research, 1978, Vol. 5, No. 4 1139-1152
© 1978

Articles

A new method for the purification and identification of covalently closed circular DNA molecules

Michael Zasloff, Gordon D. Ginder and Gary Felsenfeld

Laboratory of Molecular Biology, National Institute of Arthritis, Metabolism and Digestive Diseases, National Institutes of Health Bethesda, MD 20014, USA

Received February 2, 1978. A new technique has been developed for the rapid isolation of covalently closed circular DNA molecules. The procedure is a selective extraction based on differences in the partitioning of covalently closed circular DNA molecules and noncovalently closed species between phenol and water at acid pH and low ionic strength. Under the conditions described, linear as well as nicked circular DNA is extracted into phenol, while covalently closed circular DNA molecules remain in the water phase. The method permits the quantitative isolation of covalently closed circular DHA from either tofal cellular DNA or partially purified preparations, to a degree of purity comparable with buoyant density procedures.


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