This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by McGlynn, A. P. Articles by Downes, C. S. Search for Related Content PubMed PubMed Citation Articles by McGlynn, A. P. Articles by Downes, C. S.

The Bromodeoxyuridine Comet Assay

Detection of Maturation of Recently Replicated DNA in Individual Cells¹

Cancer and Ageing Research Group, School of Biomedical Sciences, University of Ulster, Coleraine BT52 1JD, Northern Ireland

The single-cell gel electrophoresis (Comet) assay is a relatively simple method of measuring DNA single strand breaks and alkali-labile sites in individual cells. We have combined this with bromodeoxyuridine (BrdUrd) labeling of DNA and immunolocalization of the BrdUrd to assess DNA replicative integrity on a single-cell basis. We show that the existence of strand discontinuities in recently replicated domains of DNA, caused during semiconservative replication or exacerbated by the arrest of replicative polymerases at UV irradiation- or chemical-induced lesions, can be detected in individual cells. Data obtained from BrdUrd-Comets are consistent with biochemical data derived with a range of techniques showing that DNA replication involves the creation of strand breaks or gaps adjacent to recently replicated material, and that DNA damage prolongs the duration of such discontinuities where DNA polymerases are stalled opposite lesions (R. T. Johnson et al., The Legacy of Cell Fusion, pp. 50–67, Oxford: Science Publications, 1994; R. B. Painter, J. Mol. Biol., 143: 289–301, 1980.). Compared with standard biochemical techniques, the BrdUrd-Comet assay is simple and suitable for the accurate and automatable assessment of replicative integrity in very small numbers of mammalian cells, such as may be obtained by biopsy.

This article has been cited by other articles:

				G. R. Wasson, V. J. McKelvey-Martin, and C. S. Downes The use of the comet assay in the study of human nutrition and cancer Mutagenesis, May 1, 2008; 23(3): 153 - 162. [Abstract] [Full Text] [PDF]

				H. Jiao, S. L. Allinson, M. J. Walsh, R. Hewitt, K. J. Cole, D. H. Phillips, and F. L. Martin Growth kinetics in MCF-7 cells modulate benzo[a]pyrene-induced CYP1A1 up-regulation Mutagenesis, March 1, 2007; 22(2): 111 - 116. [Abstract] [Full Text] [PDF]

				G. F. Barker, N. D. Manzo, K. L. Cotich, R. K. Shone, and A. B. Waxman DNA Damage Induced by Hyperoxia: Quantitation and Correlation with Lung Injury Am. J. Respir. Cell Mol. Biol., September 1, 2006; 35(3): 277 - 288. [Abstract] [Full Text] [PDF]

				I. Hilger, A. Rapp, K.-O. Greulich, and W. A. Kaiser Assessment of DNA Damage in Target Tumor Cells after Thermoablation in Mice Radiology, November 1, 2005; 237(2): 500 - 506. [Abstract] [Full Text] [PDF]

				V. R. Young Trace Element Biology: The Knowledge Base and its Application for the Nutrition of Individuals and Populations J. Nutr., May 1, 2003; 133(5): 1581S - 1587. [Abstract] [Full Text] [PDF]