| 摘要 |
<p><P>PROBLEM TO BE SOLVED: To provide a method for purifying plasmid DNA, by which high-purity plasmid DNA is easily mass purified in a short time. <P>SOLUTION: The method comprises the following process: Bacterial cells are suspended in a Tris-EDTA buffer, and adding a lysis buffer containing NaOH and 3-(dodecyldimethyl-ammonio)propanesulfonate salt as amphoteric surfactant to lyse the cells. Next, a Tris-buffer is added to the resultant cell lysate and centrifuged to precipitate a protein coagulation where plasmid DNA, RNA and part of proteins are loosely bound together. The resultant precipitate is washed with a Tris-buffer to remove the RNA from the protein coagulation, and a TE-buffer or water is added to the thus washed precipitate, tapped or agitated with a vortex mixer to elute the plasmid DNA from the protein coagulation. Finally, the resultant eluate is centrifuged to collect the resulting supernatant, thus obtaining the plasmid DNA. <P>COPYRIGHT: (C)2007,JPO&INPIT</p> |
