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

0580560 - ÚEM 2024 RIV US eng J - Journal Article
Collins, A. - Moller, P. - Gajski, G. - Vodenková, Soňa - Abdulwahed, A. - Anderson, A. - Bankoglu, E.E. - Bonassi, S. - Boutet-Robinet, E. - Brunborg, G. - Chao, Ch. - Cooke, M.S. - Costa, C. - Costa, S. - Dhawan, A. - de Lapuente, J. - Del Bo', C. - Dubus, J. - Dusinska, M. - Duthie, S.J. - el Yamani, N. - Engelward, B. - Gaivao, I. - Giovannelli, L. - Godschalk, R. … Total 79 authors
Measuring DNA modifications with the comet assay: a compendium of protocols.
Nature Protocols. Roč. 18, č. 3 (2023), s. 929-989. ISSN 1754-2189. E-ISSN 1750-2799
Institutional support: RVO:68378041
Keywords : cell gel-electrophoresis * double-strand breaks * in-vivo model * inter-laboratory variation * induced oxidative stress * interstrand cross-links
OECD category: Biochemistry and molecular biology
Impact factor: 14.8, year: 2022
Method of publishing: Open access
https://www.nature.com/articles/s41596-022-00754-y

The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. 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 and 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.

The comet assay is commonly used to assess DNA damage. This collection of consensus protocols includes adaptations for a wide range of species and sample types, assay formats and detection of different types of DNA lesions.
Permanent Link: https://hdl.handle.net/11104/0350866