Isolation of plasmid DNA from bacteria

Methods Enzymol. 2013:529:135-42. doi: 10.1016/B978-0-12-418687-3.00010-0.

Abstract

The purpose of this protocol is the isolation of plasmid DNA from bacteria. The boiling method for isolating plasmids by Holmes and Quigley (1981) is presented here. This method is rapid and simple and it allows for a large number of samples to be processed simultaneously (up to 40 samples). Thus, it is appropriate for the preparation of bacterial plasmids in order to screen a large number of colonies or small cultures for the presence of recombinant DNA inserts. The protocol can be effectively scaled up for the preparation of plasmids from liter cultures.

Keywords: Bacterial cells; Boiling method; Plasmid DNA; RNase; STET buffer.

MeSH terms

  • DNA, Bacterial / isolation & purification*
  • Escherichia coli / genetics
  • Plasmids / genetics
  • Plasmids / isolation & purification*

Substances

  • DNA, Bacterial