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Comet and micronucleus assays for analyzing DNA damage and genome integrity
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).ORCID iD: 0000-0003-1170-0224
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).ORCID iD: 0000-0001-7622-5116
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).ORCID iD: 0000-0002-1269-8288
2019 (English)In: DNA SENSORS AND INFLAMMASOMES / [ed] Sohn, J, ELSEVIER ACADEMIC PRESS INC , 2019, 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 ACADEMIC PRESS INC , 2019. p. 299-307
Series
Methods in Enzymology, ISSN 0076-6879 ; 625
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-164490DOI: 10.1016/bs.mie.2019.05.015ISI: 000488782900019PubMedID: 31455533ISBN: 978-0-12-818360-1 (electronic)ISBN: 978-0-12-818359-5 (print)OAI: oai:DiVA.org:umu-164490DiVA, id: diva2:1365668
Available from: 2019-10-25 Created: 2019-10-25 Last updated: 2019-10-25Bibliographically approved

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Jiang, HuiPanda, SwarupaGekara, Nelson O.

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