Modelling the comet assay

Darragh G McArt, G McKerr, Vyvyan Howard, Kurt Saetzler, Gillian R Wasson

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The single-cell gel electrophoresis technique or comet assay is widely regarded as a quick and reliable method of analysing DNA damage in individual cells. It has a proven track record from the fields of biomonitoring to nutritional studies. The assay operates by subjecting cells that are fixed in agarose to high salt and detergent lysis, thus removing all the cellular content except the DNA. By relaxing the DNA in an alkaline buffer, strands containing breaks are released from supercoiling. Upon electrophoresis, these strands are pulled out into the agarose, forming a tail which, when stained with a fluorescent dye, can be analysed by fluorescence microscopy. The intensity of this tail reflects the amount of DNA damage sustained. Despite being such an established and widely used assay, there are still many aspects of the comet assay which are not fully understood. The present review looks at how the comet assay is being used, and highlights some of its limitations. The protocol itself varies among laboratories, so results from similar studies may vary. Given such discrepancies, it would be attractive to break the assay into components to generate a mathematical model to investigate specific parameters.
LanguageEnglish
Pages914-917
JournalBiochemical Society Transactions
Volume037
Issue number4
DOIs
Publication statusPublished - 2009

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assays
DNA damage
agarose
tail
fluorescent dyes
DNA
cells
fluorescence microscopy
detergents
gel electrophoresis
electrophoresis
mathematical models
buffers
salts
methodology

Cite this

McArt, D. G., McKerr, G., Howard, V., Saetzler, K., & Wasson, G. R. (2009). Modelling the comet assay. Biochemical Society Transactions, 037(4), 914-917. https://doi.org/10.1042/BST0370914
McArt, Darragh G ; McKerr, G ; Howard, Vyvyan ; Saetzler, Kurt ; Wasson, Gillian R. / Modelling the comet assay. In: Biochemical Society Transactions. 2009 ; Vol. 037, No. 4. pp. 914-917.
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McArt, DG, McKerr, G, Howard, V, Saetzler, K & Wasson, GR 2009, 'Modelling the comet assay', Biochemical Society Transactions, vol. 037, no. 4, pp. 914-917. https://doi.org/10.1042/BST0370914

Modelling the comet assay. / McArt, Darragh G; McKerr, G; Howard, Vyvyan; Saetzler, Kurt; Wasson, Gillian R.

In: Biochemical Society Transactions, Vol. 037, No. 4, 2009, p. 914-917.

Research output: Contribution to journalArticle

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McArt DG, McKerr G, Howard V, Saetzler K, Wasson GR. Modelling the comet assay. Biochemical Society Transactions. 2009;037(4):914-917. https://doi.org/10.1042/BST0370914