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

Andrew Collins, Peter Møller, Goran Gajski, Soňa Vodenková, Abdulhadi Abdulwahed, Diana Anderson, Ezgi Eyluel Bankoglu, Stefano Bonassi, Elisa Boutet-Robinet, Gunnar Brunborg, Christy Chao, Marcus S Cooke, Carla Costa, Solange Costa, Alok Dhawan, Joaquin de Lapuente, Cristian Del Bo', Julien Dubus, Maria Dusinska, Susan J DuthieNaouale El Yamani, Bevin Engelward, Isabel Gaivão, Lisa Giovannelli, Roger Godschalk, Sofia Guilherme, Kristine B Gutzkow, Khaled Habas, Alba Hernández, Oscar Herrero, Marina Isidori, Awadhesh N Jha, Siegfried Knasmüller, Ingeborg M Kooter, Gudrun Koppen, Marcin Kruszewski, Carina Ladeira, Blanca Laffon, Marcelo Larramendy, Ludovic Le Hégarat, Angélique Lewies, Anna Lewinska, Guillermo E Liwszyc, Adela López de Cerain, Mugimane Manjanatha, Ricard Marcos, Mirta Milić, Vanessa Moraes de Andrade, Massimo Moretti, Damian Muruzabal, Ricardo Marcos Dauder

Research output: Contribution to journalReview articleResearchpeer-review

79 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)929-989
Number of pages61
JournalNature Protocols
Volume18
Issue number3
Early online date27 Jan 2023
DOIs
Publication statusPublished - 27 Jan 2023

Keywords

  • Animals
  • Comet Assay/methods
  • DNA Damage
  • DNA/genetics
  • Eukaryotic Cells
  • Humans
  • Pyrimidine Dimers

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