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Novel method for the high-throughput processing of slides for the comet assay
Open Access Publishing Fund
  • Mahsa Karbaschi, Oxidative Stress Group, University of Leicester and Department of Environmental and Occupational Health, Florida International University
  • Marcus Cooke, Oxidative Stress Group, University of Leicester and Department of Environmental and Occupational Health, Florida International University
Date of this Version
11-1-2014
Document Type
Article
Abstract

Single cell gel electrophoresis (the comet assay), continues to gain popularity as a means of assessing DNA damage. However, the assay’s low sample throughput and laborious sample workup procedure are limiting factors to its application. “Scoring”, or individually determining DNA damage levels in 50 cells per treatment, is time-consuming, but with the advent of high-throughput scoring, the limitation is now the ability to process significant numbers of comet slides. We have developed a novel method by which multiple slides may be manipulated, and undergo electrophoresis, in batches of 25 rather than individually and, importantly, retains the use of standard microscope comet slides, which are the assay convention. This decreases assay time by 60%, and benefits from an electrophoresis tank with a substantially smaller footprint, and more uniform orientation of gels during electrophoresis. Our high-throughput variant of the comet assay greatly increases the number of samples analysed, decreases assay time, number of individual slide manipulations, reagent requirements and risk of damage to slides. The compact nature of the electrophoresis tank is of particular benefit to laboratories where bench space is at a premium. This novel approach is a significant advance on the current comet assay procedure.

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Publication of this article was funded in part by Florida International University Open Access Publishing Fund.

Creative Commons License
Creative Commons Attribution 4.0
Citation Information
Mahsa Karbaschi and Marcus Cooke. "Novel method for the high-throughput processing of slides for the comet assay" (2014)
Available at: http://works.bepress.com/marcus-cooke/7/