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The Comet-FISH Assay for the Analysis of DNA Damage and Repair

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Fluorescence in situ Hybridization (FISH)

Part of the book series: Methods in Molecular Biology ((MIMB,volume 659))

Abstract

In this chapter, I describe the alkaline single-cell gel electrophoresis (Comet assay) combined with fluorescence in situ hybridization (FISH) technology, used in our laboratory, to study the incidence and repair of lesions induced in human cells by ultraviolet light. The Comet-FISH method permits the simultaneous and comparative analysis of DNA damage and its repair throughout the genome and in defined chromosomal regions. This very sensitive approach can be applied to any lesion, such as those induced by chemical carcinogens and products of cellular metabolism that can be converted to DNA single- or double-strand breaks. The unique advantages and limitations of the method for particular applications are discussed.

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Acknowledgments

The author would like to thank Alexia Chollat-Namy and Rachel A. Cox for their patience and perseverance in developing the assay, and Phil Hanawalt for believing in the project. This work was supported by a grant CA91456 from NIH.

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Authors and Affiliations

  1. Department of Biology, Stanford University, Stanford, CA, USA

    Graciela Spivak

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  1. Graciela Spivak
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Correspondence to Graciela Spivak .

Editor information

Editors and Affiliations

  1. School of Health Sciences & Social Care, Div. Biosciences, Brunel University, Kingston Lane, Uxbridge, UB83PH, United Kingdom

    Joanna M. Bridger

  2. , Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX37BN, United Kingdom

    Emanuela V. Volpi

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Spivak, G. (2010). The Comet-FISH Assay for the Analysis of DNA Damage and Repair. In: Bridger, J., Volpi, E. (eds) Fluorescence in situ Hybridization (FISH). Methods in Molecular Biology, vol 659. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-789-1_9

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  • DOI: https://doi.org/10.1007/978-1-60761-789-1_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-788-4

  • Online ISBN: 978-1-60761-789-1

  • eBook Packages: Springer Protocols

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