exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes

Detecting DNA breaks in defined regions of the genome is critical to advancing our understanding of genome stability maintenance. Here, we present exo-FISH, a protocol to label exposed single-stranded DNA in defined repetitive regions of mammalian genomes by combining in vitro restriction enzyme dig...

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Detalles Bibliográficos
Autores principales: Saayman, Xanita, Graham, Emily, Leung, Chin Wei Brian, Esashi, Fumiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10425934/
https://www.ncbi.nlm.nih.gov/pubmed/37549036
http://dx.doi.org/10.1016/j.xpro.2023.102487
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author Saayman, Xanita
Graham, Emily
Leung, Chin Wei Brian
Esashi, Fumiko
author_facet Saayman, Xanita
Graham, Emily
Leung, Chin Wei Brian
Esashi, Fumiko
author_sort Saayman, Xanita
collection PubMed
description Detecting DNA breaks in defined regions of the genome is critical to advancing our understanding of genome stability maintenance. Here, we present exo-FISH, a protocol to label exposed single-stranded DNA in defined repetitive regions of mammalian genomes by combining in vitro restriction enzyme digestion on fixed cells with fluorescence in situ hybridization (FISH). We describe steps for cell harvesting and fixation, slide treatments, and FISH probe hybridization. We then detail procedures for imaging and analysis. For complete details on the use and execution of this protocol, please refer to Saayman et al. (2023).(1)
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spelling pubmed-104259342023-08-16 exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes Saayman, Xanita Graham, Emily Leung, Chin Wei Brian Esashi, Fumiko STAR Protoc Protocol Detecting DNA breaks in defined regions of the genome is critical to advancing our understanding of genome stability maintenance. Here, we present exo-FISH, a protocol to label exposed single-stranded DNA in defined repetitive regions of mammalian genomes by combining in vitro restriction enzyme digestion on fixed cells with fluorescence in situ hybridization (FISH). We describe steps for cell harvesting and fixation, slide treatments, and FISH probe hybridization. We then detail procedures for imaging and analysis. For complete details on the use and execution of this protocol, please refer to Saayman et al. (2023).(1) Elsevier 2023-08-06 /pmc/articles/PMC10425934/ /pubmed/37549036 http://dx.doi.org/10.1016/j.xpro.2023.102487 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Saayman, Xanita
Graham, Emily
Leung, Chin Wei Brian
Esashi, Fumiko
exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes
title exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes
title_full exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes
title_fullStr exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes
title_full_unstemmed exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes
title_short exo-FISH: Protocol for detecting DNA breaks in repetitive regions of mammalian genomes
title_sort exo-fish: protocol for detecting dna breaks in repetitive regions of mammalian genomes
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10425934/
https://www.ncbi.nlm.nih.gov/pubmed/37549036
http://dx.doi.org/10.1016/j.xpro.2023.102487
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AT esashifumiko exofishprotocolfordetectingdnabreaksinrepetitiveregionsofmammaliangenomes

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