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Fast Micromethod DNA Single-Strand-Break Assay

  • Protocol
DNA Repair Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 314))

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Abstract

The Fast Micromethod is a convenient and quick fluorimetric microplate assay for the assessment of DNA single-strand breaks and their repair. This method measures the rate of unwinding of cellular DNA on exposure to alkaline conditions using a fluorescent dye which preferentially binds to double-stranded DNA, but not to single-stranded DNA or protein. The advantages of this method are that it requires only minute amounts of material (30 ng of DNA or about 3000 cells per single well), it allows simultaneous measurements of multiple samples, and it can be performed within 3 h or less (for one 96-well microplate). The Fast Micromethod can be used for the routine determination of DNA damage in cells and tissue samples after irradiation, exposure to mutagenic and carcinogenic agents, or chemotherapy.

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Notes

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    1

    Patents/patent applications: DE 19724781, DE 19933078, and PCT/EP99/01545.

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Acknowledgments

This work was supported by grants from the Commission of the European Communities (EVK3-CT-1999-00005 and ICA2-CT-2000-10047), the Bundesministerium für Bildung und Forschung (Project 0312309 and WTZUkraine/Belarus) and from Deutsche Krebshilfe (Dr. Mildred Scheel Stiftung; 10-1015-Schr 1).

Author information

Authors and Affiliations

  1. Abteilung Angewandte Molekularbiologie, Institue für Physiologische Chemie, Universität, Mainz, Germany

    Heinz C. Schröder, Oleksandra Boreiko & Werner E. G. Müller

  2. Laboratory for Marine Molecular Toxicology, Center for Marine Research, Institute Ruder Boscovic, Rovinj, Croatia

    Renato Batel

  3. BIOTECmarin GmbH, Mainz, Germany

    Heiko Schwertner

Authors
  1. Heinz C. Schröder
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  2. Renato Batel
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  3. Heiko Schwertner
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  4. Oleksandra Boreiko
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  5. Werner E. G. Müller
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Editor information

Editors and Affiliations

  1. Pharmacological Sciences, SUNY Stony Brook, Stony Brook, NY

    Daryl S. Henderson

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© 2006 Humana Press Inc.

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Schröder, H.C., Batel, R., Schwertner, H., Boreiko, O., Müller, W.E.G. (2006). Fast Micromethod DNA Single-Strand-Break Assay. In: Henderson, D.S. (eds) DNA Repair Protocols. Methods in Molecular Biology™, vol 314. Humana Press. https://doi.org/10.1385/1-59259-973-7:287

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  • DOI: https://doi.org/10.1385/1-59259-973-7:287

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-513-2

  • Online ISBN: 978-1-59259-973-8

  • eBook Packages: Springer Protocols

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