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Assays for Restriction Endonucleases Using Plasmid Substrates

  • Protocol
DNA-Protein Interactions

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 30))

Abstract

A type II restriction enzyme purchased from a commercial supplier comes with a specified number of units of enzyme activity. The units of restriction enzyme activity are defined by the minimal amount of enzyme needed to complete the digestion of 1 l.tg of bacteriophage γ DNA in 1 h. These units are usually measured by making serial dilutions of the stock solution of the enzyme, adding 1 PL from each dilution to 1 µg of phage γ DNA in a suitable buffer, incubating the reactions for 1 h at 37°C, and then analyzing the DNA by electrophoresis through agarose. This is, at best, a semiquantitative assay. It cannot yield quantitative data about the rate of the reaction of a restriction enzyme on a DNA substrate.

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Author information

Authors and Affiliations

  1. Department of Biochemistry, Centre for Molecular Recognition, University of Bristol, Bristol, UK

    Stephen E. Halford, Christian L. M. Vermote & I. Barry Vipond

  2. Department of Biochemistry, Duke University Medical Center, Durham, NC

    John D. Taylor

Authors
  1. Stephen E. Halford
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  2. John D. Taylor
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  3. Christian L. M. Vermote
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  4. I. Barry Vipond
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Editor information

Editors and Affiliations

  1. School of Biological Sciences; University of Portsmouth, Portsmouth, UK

    G. Geoff Kneale

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© 1994 Humana Press Inc.

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Halford, S.E., Taylor, J.D., Vermote, C.L.M., Barry Vipond, I. (1994). Assays for Restriction Endonucleases Using Plasmid Substrates. In: Geoff Kneale, G. (eds) DNA-Protein Interactions. Methods in Molecular Biology™, vol 30. Humana Press. https://doi.org/10.1385/0-89603-256-6:385

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  • DOI: https://doi.org/10.1385/0-89603-256-6:385

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-256-9

  • Online ISBN: 978-1-59259-517-4

  • eBook Packages: Springer Protocols

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