Abstract
This report describes a simple and efficient procedure for the isolation of plasmid DNA free from chromosomal DNA, cellular RNA, and protein. The technique comprises a modified cleared lysate procedure of D. B. Clewell and D. R. Helinski (1969, Proc. Natl. Acad. Sci. USA, 62, 1159–1166) followed by high-performance liquid chromatography on a Dupont Bioseries GF250 surface stable diol-coated silica gel permeation column (Zorbax) for the final purification of the plasmid DNA. The use of HPLC facilitates rapid and high-resolution separations within 3–4 h. Plasmid DNA produced in this manner retains its biological activity and exhibits yields equal to those obtained by the conventional cesium chloride-ethidium bromide density centrifugation method.
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