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

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  • Andrew Collins
  • Goran Gajski
  • Sona Vodenkova
  • Abdulhadi Abdulwahed
  • Diana Anderson
  • Ezgi Eyluel Bankoglu
  • Stefano Bonassi
  • Elisa Boutet-Robinet
  • Gunnar Brunborg
  • Christy Chao
  • Marcus S. S. Cooke
  • Carla Costa
  • Solange Costa
  • Alok Dhawan
  • Joaquin de Lapuente
  • Cristian Del Bo
  • Julien Dubus
  • Maria Dusinska
  • Susan J. J. Duthie
  • Naouale El Yamani
  • Bevin Engelward
  • Isabel Gaivao
  • Lisa Giovannelli
  • Roger Godschalk
  • Sofia Guilherme
  • Kristine B. B. Gutzkow
  • Khaled Habas
  • Alba Hernandez
  • Oscar Herrero
  • Marina Isidori
  • Awadhesh N. N. Jha
  • Siegfried Knasmueller
  • Ingeborg M. M. Kooter
  • Gudrun Koppen
  • Marcin Kruszewski
  • Carina Ladeira
  • Blanca Laffon
  • Marcelo Larramendy
  • Ludovic Le Hegarat
  • Angelique Lewies
  • Anna Lewinska
  • Guillermo E. E. Liwszyc
  • Adela Lopez de Cerain
  • Mugimane Manjanatha
  • Ricard Marcos
  • Mirta Milic
  • Vanessa Moraes de Andrade
  • Massimo Moretti
  • Damian Muruzabal
  • Matjaz Novak
  • Rui Oliveira
  • Ann-Karin Olsen
  • Norah Owiti
  • Mario Pacheco
  • Alok K. K. Pandey
  • Stefan Pfuhler
  • Bertrand Pourrut
  • Kerstin Reisinger
  • Emilio Rojas
  • Elise Runden-Pran
  • Julen Sanz-Serrano
  • Sergey Shaposhnikov
  • Ville Sipinen
  • Karen Smeets
  • Helga Stopper
  • Joao Paulo Teixeira
  • Vanessa Valdiglesias
  • Mahara Valverde
  • Frederique van Acker
  • Frederik-Jan van Schooten
  • Marie Vasquez
  • Johannes F. F. Wentzel
  • Maciej Wnuk
  • Annelies Wouters
  • Bojana Zegura
  • Tomas Zikmund
  • Sabine A. S. Langie
  • Amaya Azqueta

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.

Original languageEnglish
JournalNature Protocols
Pages (from-to)929–989
Number of pages61
Publication statusPublished - 2023

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