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PMID: 7009583 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

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

Journal of bacteriology ·Vol. 145 ·No. 3 ·1981-03-00 ·Pages 1365-73

Kado CI, Liu ST

Abstract

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.

MeSH Terms

Bacteriological Techniques Bacteriolysis Cloning, Molecular DNA, Bacterial/isolation & purification DNA, Recombinant/isolation & purification Enterobacteriaceae/genetics Genetic Techniques Plasmids Protein Biosynthesis Pseudomonadaceae/genetics Rhizobium/genetics

Chemicals

DNA, Bacterial DNA, Recombinant

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Kado C I

Liu S T

References (27)

27 references, click to expand

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Article Info

Journal

Journal of bacteriology

Abbr.

J Bacteriol

ISSN

0021-9193

DOI

10.1128/jb.145.3.1365-1373.1981

Published

1981-03-00

Pages

1365-73

Language

English

Region

United States

NLM ID

2985120R

PMCID

PMC217141

Subset

Grants

NCI NIH HHS · CA-11526 · United States

External Links

PubMed source DOI: 10.1128/jb.145.3.1365-1373.1981 PMC Full Text View journal details

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

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