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Scientific article
English

A method to sequence genomic sites of mitotic DNA synthesis in mammalian cells

Published inMethods in enzymology, vol. 661, p. 283-304
Publication date2021
Abstract

Under normal conditions, the genome of eukaryotic cells is faithfully replicated during S phase. However, in cells exposed to DNA polymerase inhibitors, some regions of the genome may fail to be replicated prior to mitotic entry. To prevent chromosomal breakage and loss of genomic information, mitotic DNA synthesis (MiDAS) completes replication of the genome prior to the onset of anaphase. We have developed a protocol that allows one to map the genomic regions that are replicated by MiDAS in mammalian cells. The protocol involves incorporation of a thymidine analog in nascent DNA in mitotic cells and then capture and high throughput sequencing of the nascent DNA. With this approach, sites of MiDAS can be identified at high resolution.

eng
Keywords
  • Common fragile sites
  • DNA replication
  • DNA replication stress
  • MIDAS
  • Mitotic DNA synthesis
Citation (ISO format)
MAILLER, Jonathan, PADAYACHY, Laura, HALAZONETIS, Thanos. A method to sequence genomic sites of mitotic DNA synthesis in mammalian cells. In: Methods in enzymology, 2021, vol. 661, p. 283–304. doi: 10.1016/bs.mie.2021.08.013
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ISSN of the journal0076-6879
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