A rapid method for the isolation of circular DNA using an aqueous two-phase partition system.
A method of isolating circular plasmid DNA from cleared lysates of E. coli is described. Purification is achieved by virtue of the rapid re-annealing kinetics or supercoiled DNA. After a brief denaturation step, double stranded plasmid DNA is separated from denatured chromosomal DNA and RNA in a two-phase partition system using dextran and polyethylene glycol. The method is much more rapid than the conventional dye-centrifugation technique and plasmid DNA of comparable purity and yield is obtained.
D004269:DNA, Bacterial / Q000302:isolation & purification*; D004270:DNA, Circular / Q000302:isolation & purification*; D004278:DNA, Superhelical / Q000302:isolation & purification; D004926:Escherichia coli; D008722:Methods; D009695:Nucleic Acid Renaturation; D010957:Plasmids; D012997:Solvents
D004269:DNA, Bacterial; D004270:DNA, Circular; D004278:DNA, Superhelical; D012997:Solvents