A rapid alkaline extraction procedure for screening recombinant plasmid DNA
Open accessAbstract
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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With multiple-sequencing results, some random errors can be eliminated based on plurality voting to improve sequencing accuracy.
“With the multiple-sequencing results, some random errors can be eliminated on the basis of the principle of plurality voting to improve the sequencing accuracy (Fig. 4, C and D) (51, 52).”
From Electrochemical DNA synthesis and sequencing on a single electrode with scalability for integrated data storage (2021) · cites this paper as [52]
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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
- ORCID
- Birnboim, H.C.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1979-11-24
- Volume
- 7
- Issue
- 6
- Pages
- 1513-1523
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC342324
- 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
- green
- europepmc.org
- 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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