A rapid boiling method for the preparation of bacterial plasmids

David S. Holmes, Michael Quigley

Research output: Contribution to journalArticlepeer-review

1996 Citations (Scopus)

Abstract

A simple and rapid method for preparing plasmids for restriction enzyme analysis has been developed. Bacteria are boiled for 15-40 s and an insoluble clot of genomic DNA and debris is removed by low-speed centrifugation. Plasmids are recovered from the supernatant by isopropanol precipitation and can be resuspended in buffer and immediately restricted. A 5-ml bacterial culture yields enough plasmids for many restriction enzyme digestions, but a single colony on a petri dish or a 0.5-ml miniculture will suffice for a few experiments. In addition, the procedure can be readily adapted for the preparation of plasmids from liter cultures with quantitative yields.

Original languageEnglish
Pages (from-to)193-197
Number of pages5
JournalAnalytical Biochemistry
Volume114
Issue number1
DOIs
Publication statusPublished - Jun 1981

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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