Cited by 14,686Molecular Biology Techniques and ApplicationsBacterial Genetics and BiotechnologyDNA and Nucleic Acid Chemistry

A rapid alkaline extraction procedure for screening recombinant plasmid DNA

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Birnboim, H.C., Doly, J.·1979·Nucleic Acids Research

Abstract

A procedure for extracting plasmid DNA from bacterial cells 1s described. The method 1s simple enough to permit the analysis by gel electrophoresis of 100 or more clones per day yet yields plasmid DNA which is pure enough to be digestible by restriction enzymes. The principle of the method is selective alkaline denaturation of high molecular weight chromosomal DNA while covalently closed circular DNA remains double-stranded. Adequate pH control is accomplished without using a pH meter. Upon neutralization, chromosomal DNA renatures to form an insoluble clot, leaving plasmid DNA in the supernatant. Large and small plasmid DNAs have been extracted by this method.

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Crossref · 10.1093/nar/7.6.1513

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Authors
Birnboim, H. C., & Doly, J.
Affiliations
Laboratoire de Génétique Moléculaire, Institut de Recherche en Biologie Moleculaire, F-75221 Paris Cedex 05, France
Venue
Nucleic Acids Research
Published
1979-11-24
Volume
7
Issue
6
Pages
1513-1523
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
OpenAlex
W2019410656
Cited by
14,686
References
20
Citations by year
2026: 27 · 2025: 39 · 2024: 51 · 2023: 59 · 2022: 76 · 2021: 100 · 2020: 85 · 2019: 100 · 2018: 97 · 2017: 101 · 2016: 123 · 2015: 99 · 2014: 147 · 2013: 195 · 2012: 175
FWCI
90.00
Type
journal article
Language
en
Open access
Locations
  • Nucleic Acids Research (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
Keywords
plasmid DNA extraction, alkaline extraction, recombinant plasmid, bacterial clones, DNA purification
MeSH terms
Animals, Chromosomes, Bacterial, Cloning, Molecular, DNA Restriction Enzymes, DNA, Circular, DNA, Recombinant, Escherichia coli, Methods, Molecular Weight, Plasmids, Transformation, Bacterial, Mice

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