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Molecular Toxicology Protocols pp 107–119Cite as

The Comet-FISH Technique

A Tool for Detection of Specific DNA Damage and Repair

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 291))

Abstract

The comet-FISH technique described in this protocol is a tool to detect genome region-specific DNA damage and repair. It is a combination of two established techniques, the comet assay (or single-cell gel electrophoresis, or the single-cell gel test), to separate highly fragmented from moderately or nonfragmented DNA and to measure it, and fluorescence in situ hybridization (FISH), to specifically label DNA sequences of interest. Comet-FISH exists in two versions, based on the neutral and the alkaline comet assays. A detailed description of the comet assay is given in Chapter 9, so readers who are not familiar with this technique can work directly with the protocol described here, without referring to additional protocols reported elsewhere. The neutral version of the comet assay detects double-strand breaks, while the alkaline version detects both double- and single-strand breaks as well as abasic sites or sites of incomplete repair. This chapter also details cell preparation and production of the hybridization probes adapted to the comet-FISH technique. Finally, microscopic analysis of comet-FISH results is described, and possible procedures of quantification of the specific DNA damage are presented.

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Author information

Authors and Affiliations

  1. Department for Single Cell and Single Molecule Techniques, Institute for Molecular Biotechnology Jena, Jena, Germany

    Alexander Rapp & Karl Otto Greulich

  2. Kirchhoff-Institute for Physics, University of Heidelberg, Heidelberg, Germany

    Michael Hausmann

Authors
  1. Alexander Rapp
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  2. Michael Hausmann
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  3. Karl Otto Greulich
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Editor information

Editors and Affiliations

  1. Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA

    Phouthone Keohavong  & Stephen G. Grant  & 

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© 2005 Humana Press Inc.

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Rapp, A., Hausmann, M., Greulich, K.O. (2005). The Comet-FISH Technique. In: Keohavong, P., Grant, S.G. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology™, vol 291. Humana Press. https://doi.org/10.1385/1-59259-840-4:107

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  • DOI: https://doi.org/10.1385/1-59259-840-4:107

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-084-7

  • Online ISBN: 978-1-59259-840-3

  • eBook Packages: Springer Protocols

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