Abstract
The single-cell gel electrophoresis or comet assay was initially reported in 1984 as a novel technique to directly visualize DNA damage in individual cells (1). The method was modified by adding strong alkaline conditions to denature genomic DNA and detect alkali labile lesions, greatly improving sensitivity for single-strand breaks (2,3). This simple assay has come into widespread use because of its sensitivity, need for few cells, applicability to nearly all eukaryotic cells, and ease in processing numerous samples (4).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Östling, O. and Johanson, K. J. (1984) Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem. Biophys. Res. Commun. 123, 291–298.
Olive, P. L., Banáth, J. P., and Durand, R. E. (1990) Heterogeneity in radiation-induced DNA damage and repair in tumour and normal cells measured using the “comet” assay. Radiat. Res. 122, 86–94.
Singh, N. P., McCoy, M. T., Tice, R. R., and Schneider, E. L. (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res. 175, 184–191.
Fairbairn, D. W., Olive, P. L., and O’Neill, K. L.(1995) The comet assay: a comprehensive review. Mutation Res. 339, 37–59.
Fleiss, J. L. (1981) The comparison of proportions from several independent samples: ridit analysis, in Statistical Methods for Rates and Proportions, 2nd ed. John Wiley & Sons, New York, pp. 150–157.
Klaude, M., Eriksson, S., Nygren, J., and Ahnström, G. (1996) The comet assay: mechanisms and technical considerations. Mutation Res. 363, 89–96.
Tice, R. R. (1995) The single cell/gel comet assay: a microgel electrophoretic technique for the detection of DNA damage and repair in individual cells, in Environmental Mutagenesis. (Phillips, D. H. and Venitt, S., eds.) Bios Scientific Publishers, Oxford, pp. 315–339.
Strauss, G. H. S. (1991) Non-random cell killing in cryopreservation: implications for performance of the battery of leukocyte tests (BLT), I. Toxic and immunotoxic effects. Mutat. Res. 252, 1–15.
McKelvey-Martin, V. J., Green, M. H. L., Schmezer, P., Pool-Zobel, B. L., De Méo, M. P., and Collins, A. (1993) The single cell gel electrophoresis assay (comet assay): a European review. Mutat. Res. 288, 47–63.
Huycke, M. M., Abrams, V., and Moore, D. R. (2002) Enterococcus faecalis produces extracellular superoxide and hydrogen peroxide that damages colonic epithelial cell DNA. Carcinogenesis 23, 529–536.
Huycke, M. M., Moore, D., Shepard, L., et al. (2001) Extracellular superoxide production by Enterococcus faecalis requires demethylmenaquinone and is attenuated by functional terminal quinol oxidases. Mol. Microbiol. 42, 729–740.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.,Totowa, NJ
About this protocol
Cite this protocol
Huycke, M.M. (2003). Single-Cell Gel Electrophoresis or Comet Assay of Intestinal Epithelial Cells Using Manual Scoring and Ridit Analysis. In: Hensley, K., Floyd, R.A. (eds) Methods in Biological Oxidative Stress. Methods in Pharmacology and Toxicology. Humana Press. https://doi.org/10.1385/1-59259-424-7:101
Download citation
DOI: https://doi.org/10.1385/1-59259-424-7:101
Publisher Name: Humana Press
Print ISBN: 978-0-89603-815-8
Online ISBN: 978-1-59259-424-5
eBook Packages: Springer Protocols