A rapid method for the purification of supercoiled PM2 DNA by affinity chromatography on H1 histone covalently coupled to agarose.
A simple and rapid method is described for the purification of supercoiled PM2 DNA by affinity chromatography on columns of H1 histone covalently coupled to agarose. The method does not require the use of intercalating agents or ultracentrifugation procedures. Under the conditions most appropriate for purification, elution is carried out in a single step with buffered 0.7 M NaCl after the sample has been loaded onto the column in buffered 0.2 M NaCl. The DNA eluted at the higher salt concentration consists of supercoiled closed circular DNA at greater than 90% purity independently of the ratio of supercoiled to nicked circular DNA in the input mixture.
D001435:Bacteriophages / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods; D004278:DNA, Superhelical / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification*; D006657:Histones; D011549:Pseudomonas / Q000032:analysis
D004278:DNA, Superhelical; D004279:DNA, Viral; D006657:Histones