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https://hdl.handle.net/2183/46906 Measuring DNA Modifications With the Comet Assay: A Compendium of Protocols
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Collins, Andrew
Moller, Peter
Gajski, Goran
Vodenková, Soňa
Abdulwahed, Abdulhadi
Anderson, Diana
Bankoglu, Ezgi Eyluel
Bonassi, Stefano
Boutet-Robinet, Elisa
Brunborg, Gunnar
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Collins A, Møller P, Gajski G, Vodenková S, Abdulwahed A, Anderson D, Bankoglu EE, Bonassi S, Boutet-Robinet E, Brunborg G, Chao C, Cooke MS, Costa C, Costa S, Dhawan A, de Lapuente J, Bo’ CD, Dubus J, Dusinska M, Duthie SJ, Yamani NE, Engelward B, Gaivão I, Giovannelli L, Godschalk R, Guilherme S, Gutzkow KB, Habas K, Hernández A, Herrero O, Isidori M, Jha AN, Knasmüller S, Kooter IM, Koppen G, Kruszewski M, Ladeira C, Laffon B, Larramendy M, Hégarat LL, Lewies A, Lewinska A, Liwszyc GE, de Cerain AL, Manjanatha M, Marcos R, Milić M, de Andrade VM, Moretti M, Muruzabal D, Novak M, Oliveira R, Olsen A-K, Owiti N, Pacheco M, Pandey AK, Pfuhler S, Pourrut B, Reisinger K, Rojas E, Rundén-Pran E, Sanz-Serrano J, Shaposhnikov S, Sipinen V, Smeets K, Stopper H, Teixeira JP, Valdiglesias V, Valverde M, van Acker F, van Schooten F-J, Vasquez M, Wentzel JF, Wnuk M, Wouters A, Žegura B, Zikmund T, Langie SAS, Azqueta A (2023) Measuring DNA modifications with the comet assay: a compendium of protocols. Nat Protoc 18(3):929–989. https://doi.org/10.1038/s41596-022-00754-y
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Abstract
[Abstract] 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
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This is an accepted version of the published document. This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections.