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dc.contributor.authorAnderson, Diana*
dc.contributor.authorLaubenthal, Julian*
dc.date.accessioned2016-10-27T15:50:10Z
dc.date.available2016-10-27T15:50:10Z
dc.date.issued2013
dc.identifier.citationAnderson D and Laubenthal J (2013) Analysis of DNA damage via single-cell electrophoresis. In: Makovets S (Ed) DNA Electrophoresis. (Methods in Molecular Biology) New York: Humana Press: 209-218.
dc.identifier.urihttp://hdl.handle.net/10454/10115
dc.descriptionNo
dc.description.abstractThe comet assay or single-cell gel electrophoresis assay is a relatively simple and sensitive technique for quantitatively measuring DNA damage and repair at the single-cell level in all types of tissue where a single-cell suspension can be obtained. Isolated cells are mixed with agarose, positioned on a glass slide, and then lysed in a high-salt solution which removes all cell contents except the nuclear matrix and DNA, which is finally subjected to electrophoresis. Damaged DNA is electrophoresed from the nuclear matrix into the agarose gel, resembling the appearance of a comet, while undamaged DNA remains largely within the proximity of the nuclear matrix. By choosing different pH conditions for electrophoresis, different damage types and levels of sensitivity are produced: a neutral (pH 8–9) electrophoresis mainly detects DNA double-strand breaks, while alkaline (pH ≥ 13) conditions detect double- and single-strand breaks as well as alkali-labile sites. This protocol describes a standard comet assay study for the analysis of DNA damage and outlines important variations of this protocol.
dc.subjectComet assay; Single-cell electrophoresis; DNA damage; Double-strand breaks; Single-strand breaks; Alkali-labile sites
dc.titleAnalysis of DNA damage via single-cell electrophoresis
dc.status.refereedYes
dc.typeBook chapter
dc.type.versionNo full-text available in the repository
dc.identifier.doihttps://doi.org/10.1007/978-1-62703-565-1_14


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