A simple procedure for large-scale purification of plasmid DNA.
We report a simple, rapid and reliable procedure for large-scale purification of plasmid DNA from non-amplified bacterial cultures. It is a modification of the boiling method of Holmes and Quigley [Anal. Biochem. 114 (1981) 193-197] and involves gel-filtration chromatography using Sephacryl S-1000 for final purification of plasmid DNA. This method does not require CsCl gradients and the recovered plasmids are free of RNA and chromosomal DNA, are supercoiled, retain their biological activity, and are suitable for restriction analysis.
Gómez-Márquez
J
J
Departamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Santiago, Santiago de Compostela, Spain.
Freire
M
M
Segade
F
F
D002852:Chromatography, Ion Exchange / Q000379:methods; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D010957:Plasmids
D004269:DNA, Bacterial