| 摘要 |
Closed circular DNA can be recovered from a cells or subcellular organelles containing closed circular DNA by a method comprising, in sequence, the steps of: (a) lysing the cells or organelles to release DNA, for example with a combination of lysozyme and heat or detergent; (b) enzymatically treating the released plasmid DNA with a proteolytic enzyme such as Proteinase K; (c) enzymatically treating the proteolytic enzyme-treated plasmid DNA with RNase and topoisomerase (I); (d) selectively enzymatically digesting the non-circular nucleic acids from the microorganism; and (e) recovering closed circular DNA. The method is particularly applicable to recovery and purification of plasmid DNA from E. coli. |
