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DNA interstrand cross-links (ICLs) are an extremely toxic form of DNA damage that cells experience upon exposure to natural metabolites. Moreover, ICLs are cytotoxic lesions produced by a range of clinically important anticancer agents. Therefore, improving our understanding of ICL induction and processing has important implications in 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.

Original publication

DOI

10.1007/978-1-0716-0323-9_7

Type

Chapter

Publication Date

2020

Volume

2119

Pages

79 - 88

Keywords

DNA damage, DNA repair, Electrophoresis, Fanconi anemia, Interstrand cross-links, Unhooking