An improved method for rapid purification of covalently closed circular plasmid DNA over a wide size range
- 28 June 1992
- journal article
- Published by Oxford University Press (OUP) in Letters in Applied Microbiology
- Vol. 14 (6), 250-254
- https://doi.org/10.1111/j.1472-765x.1992.tb00698.x
Abstract
An improved method has been developed for the large-scale purification of covalently closed circular (CCC) plasmid DNA molecules of sizes ranging from 4.3 to 73 kb. This protocol uses an alkaline-lysis procedure followed by acid-phenol extraction but with several modifications to previously reported methods. The principal modification is the replacement of NaCl by MgCl2 in the extraction buffer to improve yield and to remove chromosomal and other non-CCC plasmid DNA. Plasmid DNA can be purified in less than 1 h and used successfully in restriction enzyme analysis and cloning experiments.Keywords
This publication has 11 references indexed in Scilit:
- Distinct plasmid profiles of Pasteurella haemolytica serotypes and the characterization and amplification in Escherichia coli of ampicillin-resistance plasmids encoding ROB-1 -lactamaseJournal of General Microbiology, 1992
- An improved method for rapid isolation of plasmid DNA from wild-type Gram-negative bacteria for plasmid restriction profile analysisLetters in Applied Microbiology, 1990
- Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coliPlasmid, 1984
- Aspects of the mechanism of acid-phenol extraction of nucleic acidsBiochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1983
- The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primersGene, 1982
- A rapid alkaline extraction procedure for screening recombinant plasmid DNANucleic Acids Research, 1979
- Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in transProceedings of the National Academy of Sciences, 1979
- A new method for the purification and identification of covalently closed circular DNA moleculesNucleic Acids Research, 1978
- Interaction of nucleic acids: VIII. Binding of magnesium ions by nucleic acidsJournal of Molecular Biology, 1971
- A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells.Proceedings of the National Academy of Sciences, 1967