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DNA interstrand crosslinks (ICLs) are an extremely toxic form of DNA damage that cells experience upon exposure to natural metabolites. Moreover, ICLs are the cytotoxic lesion produced a range of clinically important anticancer agents. Therefore, improving our understanding of ICL induction and processing has important implications for biology and medicine. The sensitive detection of ICLs in mammalian cells is challenging but has been aided by the development of a modified form of the single-cell gel electrophoresis (SCGE) assay, also known as the 'Comet assay'. Here we describe this method and how it can be used to sensitively monitor the induction and removal of ICLs in single mammalian cells.

Type

Chapter

Book title

DNA Electrophoresis: Methods and Protocols.

Publisher

Springer Nature 2020

Publication Date

2020

Volume

2119

Addresses

Lonnie P. Swift, Weatherall Institute of Molecular Medicine, Oncology, Oxford, OX3 9DS, United Kingdom, Lianne Castle, Weatherall Institute of Molecular Medicine, Oncology, Oxford, OX3 9DS, United Kingdom, Peter J. McHugh, Weatherall Institute of Molecular Medicine, Oncology, Oxford, OX3 9DS, United Kingdom

Keywords

DNA damage, DNA repair, unhooking, interstrand crosslinks, Fanconi anaemia, electrophoresis