PURIFICATION OF PLASMID DNA HAVING SUPER-COIL FORM

摘要

PURPOSE: To purify a plasmid DNA for gene recombination use in a short time at a low cost without using harmful reagent by removing impurities from a liquid mixture of plasmid using a filter having a specific average pore diameter. ;CONSTITUTION: A plasmid DNA having super-coil form and suitable for gene recombination use is purified in a short time at a low cost without using harmful reagent by culturing E.coli transformed with a plasmid in a medium at 37°C for 1 night, collecting the microbial cells from the cultured liquid, recovering the plasmid DNA by alkali bacteriolysis and filtering the produced plasmid mixture liquid with a filter composed of a cuprammonium regenerated cellulose porous membrane hollow fiber having an average pore diameter of 10-35nm and capable of practionating DNA in a range of molecular weights of 10,000-100,000,000 under 200mmHg pressure, thereby removing the impurities from the mixture.;COPYRIGHT: (C)1996,JPO