Measuring DNA modifications with the comet assay: a compendium of protocols

The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to humans. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA-DNA crosslinks, UV-induced cyclobutane pyrimidine dimers, some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry, and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species, and types of DNA damage, thereby demonstrating its versatility.

Keywords

Comet assay DNA modifications Protocol

URI

http://hdl.handle.net/10400.21/15472

Citation

Collins A, Møller P, Gajski G, Vodenková S, Abdulwahed A, Ladeira C, et al. Measuring DNA modifications with the comet assay: a compendium of protocols. Nat Protoc. 2023;18(3):929-89.