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Mutagenesis Advance Access originally published online on February 17, 2008
Mutagenesis 2008 23(3):143-151; doi:10.1093/mutage/gem051
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© The Author 2008. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org.

The comet assay: topical issues

Andrew R. Collins1,*, Amaia Azqueta Oscoz1, Gunnar Brunborg2, Isabel Gaivão1,3, Lisa Giovannelli4, Marcin Kruszewski5, Catherine C. Smith6 and Rudolf Stetina7

1Department of Nutrition, Faculty of Medicine, University of Oslo, Oslo, Norway 2Norwegian Institute of Public Health, Oslo, Norway 3Genetic and Biotechnology Department, University of Trás-os-Montes and Alto Douro, Quinta de Prados, Vila Real, Portugal 4Department of Preclinical and Clinical Pharmacology, University of Florence, Florence, Italy 5Institute of Nuclear Chemistry and Technology, Warszawa, Poland 6AstraZeneca, Safety Assessment, R&D, Macclesfield, UK 7Faculty of Military Health Sciences, University of Defence, Hradec Kralove, Czech Republic

The comet assay is a versatile and sensitive method for measuring single- and double-strand breaks in DNA. The mechanism of formation of comets (under neutral or alkaline conditions) is best understood by analogy with nucleoids, in which relaxation of DNA supercoiling in a structural loop of DNA by a single DNA break releases that loop to extend into a halo—or, in the case of the comet assay, to be pulled towards the anode under the electrophoretic field. A consideration of the simple physics underlying electrophoresis leads to a better understanding of the assay. The sensitivity of the assay is only fully appreciated when it is calibrated: between one hundred and several thousand breaks per cell can be determined. By including lesion-specific enzymes in the assay, its range and sensitivity are greatly increased, but it is important to bear in mind that their specificity is not absolute. Different approaches to quantitation of the comet assay are discussed. Arguments are presented against trying to apply the comet assay to the study of apoptosis. Finally, some of the advantages and disadvantages of using the comet assay on lymphocyte samples collected in human studies are rehearsed.

* To whom correspondence should be addressed at Department of Nutrition, Faculty of Medicine, University of Oslo, PB 1046 Blindern, 0316 Oslo, Norway. Tel: +47 22851360; Fax: +47 22851341; Email: a.r.collins{at}medisin.uio.no

Received on October 10, 2007; revised on November 28, 2007; accepted on December 6, 2007.


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