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Title: The comet assay : a biomonitoring tool for nutraceutical research
Authors: Wong, VWC
Szeto, YT
Collins, AR
Benzie, IFF 
Issue Date: 2005
Source: Current topics in nutraceutical research, 2005, v. 3, no. 1, p. 1-14
Abstract: The comet assay is a relatively simple, but sensitive and well validated tool for measuring strand breaks in DNA in single cells. Cells are embedded in a thin layer of agarose on a microscope slide and lysed with detergent and high salt solution. This procedure also removes proteins and histones, leaving a nucleoid from each embedded cell lying within a cavity in the gel. The presence of breaks in DNA causes a local relaxation in the supercoiled loops of DNA in the nucleoid. When a small electrical charge is passed through the gel, the relaxed areas of the DNA loops are pulled towards the anode, forming a comet ‘tail’, the DNA in the nucleoid being the comet ‘head’. Comets are visualized by fluorescent microscopy, and the amount of DNA in the tail, relative to the head, is proportional to the amount of strand breaks. Cells can be incubated in vitro with an agent of interest prior to the comet assay, and the resulting DNA damage can then be measured. The effect of a ‘challenge’ to DNA after treated cells have been exposed to a putative protective agent can also be investigated, and cells (usually lymphocytes) can be collected pre- and postsupplementation with an agent of interest to assess possible genoprotective or genotoxic effects. There are various versions of the comet assay, and these make it a highly versatile biomonitoring tool for nutraceutical research. In this review, the principles and applications of the comet assay are described and its application in nutraceutical research highlighted.
Keywords: Antioxidant
Comet assay
DNA damage
Oxidative stress
Publisher: New Century Health Publishers
Journal: Current topics in nutraceutical research
ISSN: 1540-7535 (print)
Appears in Collections:Journal/Magazine Article

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