Preparation of plasmid DNA by gamma-irradiation of recA cells.
If recA bacteria are exposed to appropriate doses of gamma-irradiation, nondividing cells which can sustain the multiplication of "small" plasmids are produced. The gamma-irradiation technique has a number of advantages over other methods for preparing pure plasmid DNA: (1) there is little, if any, contamination of DNA preparations by chromosomal DNA owing to extensive degradation of the irradiated DNA by endogenous nucleases, (2) there is no need to introduce a uvr mutation to the host bacteria (there is when UV is used to inactivate the chromosome), (3) the method is extremely simple to work with since operations are not limited by considerations of volume and cell density, and (4) there is no need to transfer material from container to container. Yields of plasmid DNA obtained by the gamma-irradiation technique compare favorably with those obtained by other methods.
D003001:Cloning, Molecular / Q000379:methods*; D004260:DNA Repair / Q000528:radiation effects; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D005720:Gamma Rays; D010957:Plasmids; D011938:Rec A Recombinases / Q000235:genetics
D004269:DNA, Bacterial; D011938:Rec A Recombinases; D004262:DNA Restriction Enzymes