Rapid method for purification of plasmid DNA and DNA fragments from DNA linkers using high-performance liquid chromatography on TSK-PW gel.
High-performance size exclusion chromatography (HPSEC) using TSK-G5000 PW in Tri buffer has been found to be a reliable method for the rapid fractionation of DNA ligation products. Plasmid and fragmented phage DNAs were found to elute in less than 2 min with recoveries greater than 98%. Escherichia coli transfection studies, using plasmid DNA that had been subjected to HPSEC column fractionation, showed high transformation efficiencies. MgCl2, a component of the DNA ligation reaction, was found to produce DNA-column support interactions, which resulted in low DNA recoveries. Such interactions were eliminated by chelation with ethylenediaminetetraacetate prior to chromatography.
D001435:Bacteriophages / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli; D010957:Plasmids
D004247:DNA