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Comet and micronucleus assays for analyzing DNA damage and genome integrity
Stockholm University, Faculty of Science, Department of Molecular Biosciences, The Wenner-Gren Institute. Umeå University, Sweden.ORCID iD: 0000-0002-1269-8288
Number of Authors: 32019 (English)In: DNA Sensors and Inflammasomes / [ed] Jungsan Sohn, Elsevier, 2019, Vol. 625, p. 299-307Chapter in book (Refereed)
Abstract [en]

Detection of DNA damage in cells is fundamental for the study of DNA repair and genome-instability associated processes including carcinogenesis. Many studies often rely on cytotoxicity assays to estimate genotoxicity. However, measurements of cytotoxicity, a delayed outcome requiring high threshold genotoxicity to induce, does not provide information about the subtle, early genotoxic effects relevant for mechanistic understanding of DNA repair processes. Here describe how to combine two simple procedures for monitoring the presence of DNA damage in individual eukaryotic cells using: (1) the Comet assay for measuring initial DNA breaks and (2) the Micronucleus assay for detecting delayed outcome DNA breaks in dividing cells. We discuss the principles, experimental design considerations and troubleshooting tips for optimizing these methods. They require standard molecular biology instruments and a fluorescent microscope.

Place, publisher, year, edition, pages
Elsevier, 2019. Vol. 625, p. 299-307
Series
Methods in Enzymology, ISSN 0076-6879, E-ISSN 1557-7988 ; 625
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:su:diva-175914DOI: 10.1016/bs.mie.2019.05.015ISI: 000488782900019PubMedID: 31455533ISBN: 978-0-12-818359-5 (print)ISBN: 978-0-12-818360-1 (electronic)OAI: oai:DiVA.org:su-175914DiVA, id: diva2:1372204
Available from: 2019-11-22 Created: 2019-11-22 Last updated: 2019-12-04Bibliographically approved

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Jiang, HuiGekara, Nelson O.
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