A method for the purification of E. coli plasmid DNA by homogeneous lysis and polyethylene glycol precipitation.

Molecular biology reports ·Vol. 9 ·No. 3 ·1983-08-00 ·Pages 191-5

Pulleyblank D, Michalak M, Daisley SL, Glick R

Abstract

A procedure is described for the isolation and purification of E. coli plasmid DNA by polyethylene glycol precipitation. The method is rapid, simple, inexpensive and amenable to both small and large scale manipulation. This procedure involves lysis of bacterial cells by treatment with pronase in sodium dodecyl sulfate, removal of chromosomal DNA by centrifugation, precipitation of residual nucleic acids with polyethylene glycol and removal of RNA by precipitation with LiCl. Plasmid DNA purified as described is pure enough for restriction endonuclease analysis, for use as a vector for the cloning of cDNA or synthetic DNA, or for use as a template in an E. coli transcription-translation cell-free system.

MeSH Terms

DNA/isolation & purification Electrophoresis, Agar Gel Escherichia coli/genetics Methods Plasmids Polyethylene Glycols

Chemicals

Polyethylene Glycols DNA

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Pulleyblank D

Michalak M

Daisley S L

Glick R

References (16)

16 references, click to expand

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Article Info

Journal

Molecular biology reports

Abbr.

Mol Biol Rep

ISSN

0301-4851

DOI

10.1007/BF00775367

Published

1983-08-00

Pages

191-5

Language

English

Region

Netherlands

NLM ID

0403234

Subset

External Links

PubMed source DOI: 10.1007/BF00775367 View journal details

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A method for the purification of E. coli plasmid DNA by homogeneous lysis and polyethylene glycol precipitation.

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