Alkaline Comet Assay to Detect DNA Damage

Kade D Walsh; Takamitsu A Kato

doi:10.1007/978-1-0716-2433-3_7

Alkaline Comet Assay to Detect DNA Damage

Methods Mol Biol. 2023:2519:65-72. doi: 10.1007/978-1-0716-2433-3_7.

Authors

Kade D Walsh¹, Takamitsu A Kato²

Affiliations

¹ Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA.
² Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA. Takamitsu.Kato@colostate.edu.

PMID: 36066710
DOI: 10.1007/978-1-0716-2433-3_7

Abstract

The comet assay is an effective method for identifying DNA breaks and alkali-labile sites induced by genotoxins. Performed as a single-cell electrophoresis, this assay is especially simplistic, and the results are easily reproducible. DNA breakage can be quantitatively assessed by the induced comet tail regions, which can be measured using a variety of comet software. This protocol will finish within approximately two hours with adequate preparation, and digitized images can be taken using a confocal or standard fluorescence microscopes after staining the cell nucleus with a DNA dye.

Keywords: Alkaline lysis; Comet assay; Single-cell electrophoresis.

MeSH terms

Comet Assay / methods
DNA
DNA Damage*
Mutagens*
Staining and Labeling

Substances

Mutagens
DNA