Measurement of oxidatively-induced clustered DNA lesions using a novel adaptation of single cell gel electrophoresis (comet assay)

Georgakilas, AG; Holt, SM; Hair, JM; Loftin, CW

dc.contributor.author	Georgakilas, AG	en
dc.contributor.author	Holt, SM	en
dc.contributor.author	Hair, JM	en
dc.contributor.author	Loftin, CW	en
dc.date.accessioned	2014-03-01T01:59:44Z
dc.date.available	2014-03-01T01:59:44Z
dc.date.issued	2010	en
dc.identifier.issn	19342500	en
dc.identifier.uri	https://dspace.lib.ntua.gr/xmlui/handle/123456789/29033
dc.subject	Complex DNA damage	en
dc.subject	Oxidatively-induced clustered DNA lesions	en
dc.subject	Repair enzymes	en
dc.subject	Single-cell gel electrophoresis	en
dc.subject.other	agar gel electrophoresis	en
dc.subject.other	article	en
dc.subject.other	comet assay	en
dc.subject.other	DNA damage	en
dc.subject.other	double stranded DNA break	en
dc.subject.other	ionizing radiation	en
dc.subject.other	oxidative stress	en
dc.subject.other	priority journal	en
dc.subject.other	pulsed field gel electrophoresis	en
dc.subject.other	single stranded DNA break	en
dc.title	Measurement of oxidatively-induced clustered DNA lesions using a novel adaptation of single cell gel electrophoresis (comet assay)	en
heal.type	journalArticle	en
heal.identifier.primary	10.1002/0471143030.cb0611s49	en
heal.identifier.secondary	http://dx.doi.org/10.1002/0471143030.cb0611s49	en
heal.identifier.secondary	6.11	en
heal.publicationDate	2010	en
heal.abstract	The two basic groups of complex DNA damage are double-strand breaks (DSBs) and non-DSB oxidatively-induced clustered DNA lesions (OCDLs). The single-cell gel electrophoresis (SCGE) or comet assay has been widely used for the detection of low levels of various types of DNA lesions including single-strand breaks (SSBs), DSBs, and oxidized bases per individual cell. There are limited data on the use of the comet assay for the detection of non-DSB clustered DNA lesions using different repair enzymes as enzymatic probes. This unit discusses a novel adaptation of the comet assay used to measure these unique types of lesions. Until now OCDL yields have been measured using primarily pulsed-field agarose gel electrophoresis. The advantages offered by the current approach are: (1) measurement of OCDL levels per individual cell; (2) use of a small number of cells (~10,000) and relatively low doses of ionizing radiation (1 to 2 Gy) or low levels of oxidative stress, which are not compatible with standard agarose gel electrophoresis; and finally, (3) the assay is fast and allows direct comparison with pulsed-field gel electrophoresis results. Curr. Protoc. Cell Biol. 49:6.11.1-6.11.17. © 2010 by John Wiley & Sons, Inc.	en
heal.journalName	Current Protocols in Cell Biology	en
dc.identifier.doi	10.1002/0471143030.cb0611s49	en
dc.identifier.issue	SUPPL.49	en