Measuring DNA modifications with the comet assay: a compendium of protocols.
Journal
Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307
Informations de publication
Date de publication:
03 2023
03 2023
Historique:
received:
16
07
2021
accepted:
05
07
2022
pubmed:
29
1
2023
medline:
14
3
2023
entrez:
28
1
2023
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 36707722
doi: 10.1038/s41596-022-00754-y
pii: 10.1038/s41596-022-00754-y
pmc: PMC10281087
mid: NIHMS1896856
doi:
Substances chimiques
Pyrimidine Dimers
0
DNA
9007-49-2
Types de publication
Journal Article
Review
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
929-989Subventions
Organisme : NIEHS NIH HHS
ID : P42 ES027707
Pays : United States
Organisme : NIEHS NIH HHS
ID : R41 ES030274
Pays : United States
Informations de copyright
© 2023. Springer Nature Limited.
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