A method to sequence genomic sites of mitotic DNA synthesis in mammalian cells.
Methods Enzymol
; 661: 283-304, 2021.
Article
em En
| MEDLINE
| ID: mdl-34776216
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Replicação do DNA
/
Mitose
Limite:
Animals
Idioma:
En
Revista:
Methods Enzymol
Ano de publicação:
2021
Tipo de documento:
Article
País de afiliação:
Suíça