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
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author Moore, Gemma
Jimenez Sainz, Judit
Jensen, Ryan B.
author_facet Moore, Gemma
Jimenez Sainz, Judit
Jensen, Ryan B.
author_sort Moore, Gemma
collection PubMed
description 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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spelling pubmed-90929942022-05-12 DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells Moore, Gemma Jimenez Sainz, Judit Jensen, Ryan B. STAR Protoc Protocol 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. Elsevier 2022-05-05 /pmc/articles/PMC9092994/ /pubmed/35573479 http://dx.doi.org/10.1016/j.xpro.2022.101371 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Moore, Gemma
Jimenez Sainz, Judit
Jensen, Ryan B.
DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells
title DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells
title_full DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells
title_fullStr DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells
title_full_unstemmed DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells
title_short DNA fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells
title_sort dna fiber combing protocol using in-house reagents and coverslips to analyze replication fork dynamics in mammalian cells
topic Protocol
url 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
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