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Measuring DNA modifications with the comet assay: a compendium of protocols
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SYSNO ASEP 0580560 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Measuring DNA modifications with the comet assay: a compendium of protocols Author(s) Collins, A. (NO)
Moller, P. (DK)
Gajski, G. (HR)
Vodenková, Soňa (UEM-P) ORCID, RID
Abdulwahed, A. (US)
Anderson, A. (GB)
Bankoglu, E.E. (DE)
Bonassi, S. (IT)
Boutet-Robinet, E. (FR)
Brunborg, G. (NO)
Chao, Ch. (US)
Cooke, M.S. (US)
Costa, C. (PT)
Costa, S. (PT)
Dhawan, A. (IN)
de Lapuente, J. (ES)
Del Bo', C. (IT)
Dubus, J. (FR)
Dusinska, M. (NO)
Duthie, S.J. (GB)
el Yamani, N. (NO)
Engelward, B. (US)
Gaivao, I. (PT)
Giovannelli, L. (GB)
Godschalk, R. (NL)Number of authors 79 Source Title Nature Protocols. - : Nature Publishing Group - ISSN 1754-2189
Roč. 18, č. 3 (2023), s. 929-989Number of pages 61 s. Language eng - English Country US - United States Keywords cell gel-electrophoresis ; double-strand breaks ; in-vivo model ; inter-laboratory variation ; induced oxidative stress ; interstrand cross-links OECD category Biochemistry and molecular biology Method of publishing Open access Institutional support UEM-P - RVO:68378041 UT WOS 000921193000001 EID SCOPUS 85146929563 DOI 10.1038/s41596-022-00754-y Annotation 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.Workplace Institute of Experimental Medicine Contact Lenka Koželská, lenka.kozelska@iem.cas.cz, Tel.: 241 062 218, 296 442 218 Year of Publishing 2024 Electronic address https://www.nature.com/articles/s41596-022-00754-y
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