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The Comet Assay: A Sensitive Genotoxicity Test for the Detection of DNA Damage and Repair

2012; Springer Science+Business Media; Linguagem: Inglês

10.1007/978-1-61779-998-3_6

1940-6029

Günter Speit, Andreas Rothfuß,

Genetically Modified Organisms Research

The comet assay (single-cell gel electrophoresis) is a simple and sensitive method for studying DNA damage and repair. In this microgel electrophoresis technique, a small number of cells suspended in a thin agarose gel on a microscope slide is lysed, electrophoresed, and stained with a fluorescent DNA-binding dye. Cells with increased DNA damage display increased migration of chromosomal DNA from the nucleus toward the anode, which resembles the shape of a comet. The assay has manifold applications in fundamental research for DNA damage and repair, in genotoxicity testing of novel chemicals and pharmaceuticals, environmental biomonitoring, and human population monitoring. This chapter describes a standard protocol of the alkaline comet assay and points to some useful modifications.