• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Mar 1981; 145(3): 1365–1373.
PMCID: PMC217141

Rapid procedure for detection and isolation of large and small plasmids.


Procedures are described for the detection and isolation of plasmids of various sizes (2.6 to 350 megadaltons) that are harbored in species of Agrobacterium, Rhizobium, Escherichia, Salmonella, Erwinia, Pseudomonas, and Xanthomonas. The method utilized the molecular characteristics of covalently closed circular deoxyribonucleic acid (DNA) that is released from cells under conditions that denature chromosomal DNA by using alkaline sodium dodecyl sulfate (pH 12.6) at elevated temperatures. Proteins and cell debris were removed by extraction with phenol-chloroform. Under these conditions chromosomal DNA concentrations were reduced or eliminated. The clarified extract was used directly for electrophoretic analysis. These procedures also permitted the selective isolation of plasmid DNA that can be used directly in nick translation, restriction endonuclease analysis, transformation, and DNA cloning experiments.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barnes WM. Plasmid detection and sizing in single colony lysates. Science. 1977 Jan 28;195(4276):393–394. [PubMed]
  • Bazaral M, Helinski DR. Circular DNA forms of colicinogenic factors E1, E2 and E3 from Escherichia coli. J Mol Biol. 1968 Sep 14;36(2):185–194. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Blin N, von Gabain A, Bujard H. Isolation of large molecular weight DNA from agarose gels for further digestion by restriction enzymes. FEBS Lett. 1975 Apr 15;53(1):84–86. [PubMed]
  • Chapman RE, Jr, Sturtevant JM. Volume changes accompanying thermal denaturation of deoxyribonucleic acid. II. Denaturation at alkaline pH. Biopolymers. 1970;9(4):445–447. [PubMed]
  • Curiale MS, Mills D. Detection and characterization of plasmids in Pseudomonas glycinea. J Bacteriol. 1977 Jul;131(1):224–228. [PMC free article] [PubMed]
  • Currier TC, Nester EW. Isolation of covalently closed circular DNA of high molecular weight from bacteria. Anal Biochem. 1976 Dec;76(2):431–441. [PubMed]
  • Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. [PubMed]
  • Gantotti BV, Patil SS, Mandel M. Apparent Involvement of a Plasmid in Phaseotoxin Production by Pseudomonas phaseolicola. Appl Environ Microbiol. 1979 Mar;37(3):511–516. [PMC free article] [PubMed]
  • Guerry P, LeBlanc DJ, Falkow S. General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973 Nov;116(2):1064–1066. [PMC free article] [PubMed]
  • Hansen JB, Olsen RH. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. [PMC free article] [PubMed]
  • Klein RD, Selsing E, Wells RD. A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid. 1980 Jan;3(1):88–91. [PubMed]
  • Lippincott BB, Margot JB, Lippincott JA. Plasmid content and tumor initiation complementation by Agrobacterium tumefaciens IIBNV6. J Bacteriol. 1977 Dec;132(3):824–831. [PMC free article] [PubMed]
  • Lurquin PF, Kado CI. Escherichia coli plasmid pBR313 insertion into plant protoplasts and into their nuclei. Mol Gen Genet. 1977 Jul 20;154(2):113–121. [PubMed]
  • Manis JJ, Whitfield HJ. Physical characterization of a plasmid cointegrate containing an F'his gnd element and the Salmonella typhimurium LT2 cryptic plasmid. J Bacteriol. 1977 Mar;129(3):1601–1606. [PMC free article] [PubMed]
  • McDonell MW, Simon MN, Studier FW. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. [PubMed]
  • Meyers JA, Sanchez D, Elwell LP, Falkow S. Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J Bacteriol. 1976 Sep;127(3):1529–1537. [PMC free article] [PubMed]
  • Mickel S, Arena V, Jr, Bauer W. Physical properties and gel electrophoresis behavior of R12-derived plasmid DNAs. Nucleic Acids Res. 1977;4(5):1465–1482. [PMC free article] [PubMed]
  • Pemberton JM, Clark AJ. Detection and characterization of plasmids in Pseudomonas aeruginosa strain PAO. J Bacteriol. 1973 Apr;114(1):424–433. [PMC free article] [PubMed]
  • Radloff R, Bauer W, Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. [PMC free article] [PubMed]
  • Record MT., Jr Electrostatic effects on polynucleotide transitions. II. Behavior of titrated systems. Biopolymers. 1967;5(10):993–1008. [PubMed]
  • RUSHIZKY GW, GRECO AE, HARTLEY RW, Jr, SOBER HA. Concentration and desalting of ribonucleases. Biochem Biophys Res Commun. 1963 Feb 18;10:311–314. [PubMed]
  • Telford J, Boseley P, Schaffner W, Birnstiel M. Novel screening procedure for recombinant plasmids. Science. 1977 Jan 28;195(4276):391–393. [PubMed]
  • Thuring RW, Sanders JP, Borst P. A freeze-squeeze method for recovering long DNA from agarose gels. Anal Biochem. 1975 May 26;66(1):213–220. [PubMed]
  • Tshitenge G, Luyindula N, Lurquin PF, Ledoux L. Plasmid deoxyribonucleic acid in Rhizobium vigna and Rhizobium trifolii. Biochim Biophys Acta. 1975 Dec 19;414(3):357–361. [PubMed]
  • Willshaw GA, Smith HR, Anderson ES. Application of agarose gel electrophoresis to the characterization of plasmid DNA in drug-resistant enterobacteria. J Gen Microbiol. 1979 Sep;114(1):15–25. [PubMed]
  • Zimmer C. Alkaline denaturation of DNA's from various sources. Biochim Biophys Acta. 1968 Jul 23;161(2):584–586. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...