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A rapid boiling method for the preparation of bacterial plasmids

✍ Scribed by David S. Holmes; Michael Quigley

Publisher
Elsevier Science
Year
1981
Tongue
English
Weight
1005 KB
Volume
114
Category
Article
ISSN
0003-2697

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✦ Synopsis

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.

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