Purification of supercoiled plasmids from crude cell lysates using high performance anion exchange chromatography.
High performance liquid chromatography is of increasing importance in the purification of nucleic acids. Recently, a new anion exchange column called Gen-Pak FAX has been introduced for this purpose. Previously, it has been used in the purification of restriction fragments and oligonucleotides. In this paper we present the use of the Gen-Pak FAX column for the purification of plasmids from crude E. coli lysates. The different conformational forms of the plasmid can be well separated and collected with high recoveries of both mass and activity. Up to 50 micrograms of supercoiled plasmid can be purified in a single 30 min run with up to 98% purity.
Merion
M
M
Waters Chromatography Division, Millipore Corporation, Milford, MA 01757.
Warren
W
W
D001709:Biotechnology; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D002852:Chromatography, Ion Exchange / Q000295:instrumentation*; D004269:DNA, Bacterial / Q000302:isolation & purification / Q000648:ultrastructure; D004278:DNA, Superhelical / Q000302:isolation & purification* / Q000648:ultrastructure; D004926:Escherichia coli / Q000032:analysis; D010957:Plasmids
D004269:DNA, Bacterial; D004278:DNA, Superhelical