title: Alkaline Comet Assay for Assessing DNA Damage in Individual Cells.
abstract: Single-cell gel electrophoresis, commonly called a comet assay, is a simple and sensitive method for assessing DNA damage at the single-cell level. It is an important technique in genetic toxicological studies. The comet assay performed under alkaline conditions (pH >13) is considered the optimal version for identifying agents with genotoxic activity. The alkaline comet assay is capable of detecting DNA double-strand breaks, single-strand breaks, alkali-labile sites, DNA-DNA/DNA-protein cross-linking, and incomplete excision repair sites. The inclusion of digestion of lesion-specific DNA repair enzymes in the procedure allows the detection of various DNA base alterations, such as oxidative base damage. This unit describes alkaline comet assay procedures for assessing DNA strand breaks and oxidative base alterations. These methods can be applied in a variety of cells from in vitro and in vivo experiments, as well as human studies.
authors.affiliation: Biomolecular Research Center, Boise State University, Boise, Idaho.
authors.forename: Xinzhu
authors.initials: X
authors.lastname: Pu
authors.affiliation: Department of Environmental Health, Indiana University, Bloomington, Indiana.
authors.forename: Zemin
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Environmental Health, Indiana University, Bloomington, Indiana.
authors.forename: James E
authors.initials: JE
authors.lastname: Klaunig
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D020552:Comet Assay / Q000379:methods*; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D015386:Hazardous Substances; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects*; D018384:Oxidative Stress; D012680:Sensitivity and Specificity
chemical_list: D015386:Hazardous Substances
keywords: DNA strand break; comet assay; genotoxicity; oxidative DNA damage