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The use of cetyltrimethylammonium bromide for the rapid isolation from Staphylococcus aureus of relaxable and non‐relaxable plasmid DNA suitable for in vitro manipulation
Author(s) -
Townsend D. E.,
Ashdown N.,
Bolton S.,
Grubb W. B.
Publication year - 1985
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.1985.tb01496.x
Subject(s) - plasmid , plasmid preparation , dna , restriction enzyme , biology , staphylococcus aureus , bacteria , transformation (genetics) , microbiology and biotechnology , chemistry , biochemistry , genetics , gene , pbr322
Plasmid DNA was isolated from Staphylococcus aureus by a rapid method that depends on the precipitation of DNA from cleared lysates by cetyltrimethylammonium bromide at low salt concentrations. The method was validated by its ability to provide DNA for restriction analysis of a highly relaxable plasmid species that is not isolated by more traditional techniques. The DNA can be digested with restriction endonucleases and used for transformation without further purification. The method also provides the basis for analysing staphylococcal plasmids that display a high frequency of deletion after transfer. Simple modifications of the technique allow plasmid DNA to be isolated from other bacteria and the rapid purification of DNA samples before in vitro manipulation.