Abstract
There are a number of methods available for screening either potential recombinant clones or natural isolates for the possession of plasmids. The method described in this chapter is based on that described by Sherratt (1) and is the simplest and most rapid of these. The name arises from the fact that the technique was devised to yield results using a single colony that can be taken directly from an appropriate agar plate. Although this procedure works best for high copy number plasmids, it can also be used to detect plasmids in environmental isolates, for example.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sherratt, D. (1979) Plasmid vectors for genetic manipulation in vitro. In Biochemistry of Genetic Engineering (Garland, P B and Williamson, R , eds.), pp. 29–36. The Biochemical Society, London. (Biochemical Society Symposium No. 44).
Bidwell, J. L., Lewis, D. A., and Reeves, D. S. (1981) A rapid single colony lysate method for the selective visualisation of plasmids in Enterobacteriaceae, including Serratia marcescens. J. Antimicrob. Chemother. 8, 481–485.
Birnboim, H. C, and Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7, 1513–1523.
Holmes, D. S., and Quigley, M. (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal. Biochem. 114, 193–197.
Barnes, W. M. (1977) Plasmid detection and sizing in single colony lysates. Science 195, 393–394.
Author information
Authors and Affiliations
Editor information
Rights and permissions
Copyright information
© 1984 The Humana Press Inc.
About this protocol
Cite this protocol
Dale, J.W., Greenaway, P.J. (1984). Plasmid Screening Using Single Colony Lysates. In: Walker, J.M. (eds) Nucleic Acids. Methods in Molecular Biology, vol 2. Humana Press. https://doi.org/10.1385/0-89603-064-4:289
Download citation
DOI: https://doi.org/10.1385/0-89603-064-4:289
Publisher Name: Humana Press
Print ISBN: 978-0-89603-064-0
Online ISBN: 978-1-59259-489-4
eBook Packages: Springer Protocols