Method for RNase- and organic solvent-free plasmid DNA purification using tangential flow filtration

摘要

A process for purifying plasmid DNA from prokaryotic cells comprising: (a) digesting the cells; (b) incubating the cells in the presence of alkali and a detergent for about 4 to 24 hours, to effect lysis and solubilisation thereof; (c) removing lysate contaminants from the cells to provide a plasmid DNA solution; (d) filtering the solution through a tangential flow filtration device to obtain a retentate containing the plasmid DNA; and (e) collecting the retentate whereby enzymes are not used in any of the above steps to digest RNA.