Journal Article

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

Vodenková, Soňa, Pacheco, Mário, Massachusetts Institute of Technology (MIT), Møller, Peter, Pandey, Alok, Laffon, Blanca, Lewies, Angélique, Owiti, Norah, Rundén-Pran, Elise, Centre for Environmental and Marine Studies [Aveiro] (CESAM) ; Universidade de Aveiro = University of Aveiro, Valverde, Mahara, Teixeira, João Paulo, Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele [Roma] (IRCCS San Raffaele Pisana), Zikmund, Tomas, Godschalk, Roger, Duthie, Susan, van Acker, Frederique, Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM) ; Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)) ; Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Bankoglu, Ezgi Eyluel, Università degli Studi di Milano = University of Milan (UNIMI), Department of Biological Engineering [Cambridge] (BE) ; Massachusetts Institute of Technology (MIT

2023

Overview

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.

• Publisher: Nature Publishing Group,CCSD,Nature Publishing Group UK
• Subject: 631/208/211 / 631/337/1427 / 692/53/2423 / 704/172 / Adaptation / Adducts / Alkylation / Analytical Chemistry / Animals / Apyrimidinic sites / Bases (nucleic acids) / Bioassays / Biological Techniques / Biomedical and Life Sciences / Biomedical materials / Comet assay / Comet Assay - methods / Computational Biology/Bioinformatics / Cyclobutane / Cyclobutane pyrimidine dimers / Damage assessment / Damage detection / Deoxyribonucleic acid / DNA / DNA - genetics / DNA adducts / DNA Damage / Eukaryotic Cells / Genotoxicity / Humans / In vivo methods and tests / Life Sciences / Microarrays / Organic Chemistry / Protocol / Pyrimidine Dimers / Yeast
• ISBN: 0009211930000