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.
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Conference Article|
July 22 2009
Modelling the comet assay
Darragh G. McArt
;
Darragh G. McArt
1
1
Biomedical Sciences Research Institute, University of Ulster, Coleraine, Co. Londonderry BT52 1SA, Northern Ireland, U.K.1
To whom correspondence should be addressed (email mcart-d@email.ulster.ac.uk).
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George McKerr
;
George McKerr
1
Biomedical Sciences Research Institute, University of Ulster, Coleraine, Co. Londonderry BT52 1SA, Northern Ireland, U.K.
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C. Vyvyan Howard
;
C. Vyvyan Howard
1
Biomedical Sciences Research Institute, University of Ulster, Coleraine, Co. Londonderry BT52 1SA, Northern Ireland, U.K.
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Kurt Saetzler
;
Kurt Saetzler
1
Biomedical Sciences Research Institute, University of Ulster, Coleraine, Co. Londonderry BT52 1SA, Northern Ireland, U.K.
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Gillian R. Wasson
Gillian R. Wasson
1
Biomedical Sciences Research Institute, University of Ulster, Coleraine, Co. Londonderry BT52 1SA, Northern Ireland, U.K.
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Biochem Soc Trans (2009) 37 (4): 914-917.
Article history
Received:
December 09 2008
Citation
Darragh G. McArt, George McKerr, C. Vyvyan Howard, Kurt Saetzler, Gillian R. Wasson; Modelling the comet assay. Biochem Soc Trans 1 August 2009; 37 (4): 914–917. doi: https://doi.org/10.1042/BST0370914
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