Work

Single-molecule Analysis of DNA Replication Dynamics in Budding Yeast and Human Cells by DNA Combing

Year: 2017

Type: article

Abstract: The DNA combing method allows the analysis of DNA replication at the level of individual DNA molecules stretched along silane-coated glass coverslips. Before DNA extraction, ongoing DNA synthesis is labeled with halogenated analogues of thymidine. Replication tracks are visualized by immunofluorescence using specific antibodies. Unlike biochemical and NGS-based methods, DNA combing provides unique information on cell-to-cell variations in DNA replication profiles, including initiation and elongation. Finally, this assay can be used to monitor the effect of DNA lesions on fork progression, arrest and restart.

Source: BIO-PROTOCOL

DOI: 10.21769/bioprotoc.2305

Authors: Hélène Tourrière, Julie Saksouk, Armelle Lengronne, Philippe Pasero

Institutions: Centre National de la Recherche Scientifique, Université de Montpellier, Institut de Génétique Humaine

Language: English

Topic: DNA Repair Mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: Agence Nationale de la Recherche, Ligue Contre le Cancer, Institut National Du Cancer

OpenAlex ID: W2622532511

FWCI: 0.435

Cites: 22

Cited by: 11

Related to: 10