Large-scale isolation of plasmid DNA using cetyltrimethylammonium bromide.
A rapid procedure for the large-scale isolation of plasmid DNA is described. The method utilizes cetyltrimethylammonium bromide to precipitate the plasmid following extraction of DNA by lysozyme digestion and boiling. The plasmid is then purified by passing through the spin column pZ523. The purity and yield of the plasmid obtained with this method is similar to that isolated by cesium chloride-ethidium bromide gradient centrifugation. The method does not involve any phenol-chloroform extractions and takes five to six hours for completion after growth of the bacterial cells. The plasmid obtained is amenable to digestion with various restriction endonucleases, can be used for cloning with high efficiency and is also suitable as template for dideoxy sequencing.
Ishaq
M
M
Dept. of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia 19104-6049.
Wolf
B
B
Ritter
C
C
D002499:Centrifugation, Density Gradient; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D011232:Chemical Precipitation; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004274:DNA, Recombinant / Q000302:isolation & purification*; D004926:Escherichia coli / Q000032:analysis; D010957:Plasmids
D002593:Cetrimonium Compounds; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004262:DNA Restriction Enzymes; D000077286:Cetrimonium