DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells

DNA fiber combing is a versatile technique that provides insight into replication fork dynamics at single-molecule resolution. DNA fibers are bound to silanized coverslips and combed, which straightens and aligns the fibers along a single axis. Here, we present a DNA fiber combing protocol that does...

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Detalles Bibliográficos
Autores principales: Moore, Gemma, Jimenez Sainz, Judit, Jensen, Ryan B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9092994/
https://www.ncbi.nlm.nih.gov/pubmed/35573479
http://dx.doi.org/10.1016/j.xpro.2022.101371
Descripción
Sumario:DNA fiber combing is a versatile technique that provides insight into replication fork dynamics at single-molecule resolution. DNA fibers are bound to silanized coverslips and combed, which straightens and aligns the fibers along a single axis. Here, we present a DNA fiber combing protocol that does not use commercial kits; we detail the steps to prepare all materials, reagents, and silanized coverslips. We describe the use of DLD-1 cells, but the protocol is amenable to other cell types.

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