Rapid purification of bacterial plasmids and coliphage M13 RF without CsCl centrifugation

D. S. Holmes

doi:10.1016/0003-2697(82)90198-1

Rapid purification of bacterial plasmids and coliphage M13 RF without CsCl centrifugation

D. S. Holmes

Life Sciences School

Research output: Contribution to journal › Article › peer-review

7 Citations (Scopus)

Abstract

A simple and rapid procedure for the purification of plasmids from Escherichia coli K12 has been developed. Bacterial cells are subjected to the boiling procedure [D. S. Holmes, and M. Quigley Anal. Biochem. 114, 193-197 (1981)] followed by removal of contaminating RNA by chromatography on Sepharose 2B and of genomic DNA by acid-phenol extraction. Plasmids are recovered with good yield. They can be restricted and ligated and will transform host cells. A simple modification of the procedure allows it to be used for the isolation of coliphage M13 RF DNA.

Original language	English
Pages (from-to)	428-433
Number of pages	6
Journal	Analytical Biochemistry
Volume	127
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(82)90198-1
Publication status	Published - 1 Jan 1982

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0003-2697(82)90198-1

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AB - A simple and rapid procedure for the purification of plasmids from Escherichia coli K12 has been developed. Bacterial cells are subjected to the boiling procedure [D. S. Holmes, and M. Quigley Anal. Biochem. 114, 193-197 (1981)] followed by removal of contaminating RNA by chromatography on Sepharose 2B and of genomic DNA by acid-phenol extraction. Plasmids are recovered with good yield. They can be restricted and ligated and will transform host cells. A simple modification of the procedure allows it to be used for the isolation of coliphage M13 RF DNA.

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Rapid purification of bacterial plasmids and coliphage M13 RF without CsCl centrifugation

Abstract

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