Protocol for analysis of G2/M DNA synthesis in human cells

G2/M DNA synthesis (G-MiDS) can be observed in one in five G2/M cells in unperturbed conditions by immunofluorescence microscopy. However, little is known of the genomic sites undergoing G-MiDS. Here, we describe a protocol which allows enriching for G2/M cells and investigating the sites of G-MiDS...

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Detalles Bibliográficos
Autores principales: Wang, Jianming, Saponaro, Marco
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8178111/
https://www.ncbi.nlm.nih.gov/pubmed/34136835
http://dx.doi.org/10.1016/j.xpro.2021.100570
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author Wang, Jianming
Saponaro, Marco
author_facet Wang, Jianming
Saponaro, Marco
author_sort Wang, Jianming
collection PubMed
description G2/M DNA synthesis (G-MiDS) can be observed in one in five G2/M cells in unperturbed conditions by immunofluorescence microscopy. However, little is known of the genomic sites undergoing G-MiDS. Here, we describe a protocol which allows enriching for G2/M cells and investigating the sites of G-MiDS using BrdU-seq. This method can also be used to study the role of DNA replication or transcription-associated factors in affecting G-MiDS levels in different cell lines. For complete details on the use and execution of this protocol, please refer to Wang et al. (2021).
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spelling pubmed-81781112021-06-15 Protocol for analysis of G2/M DNA synthesis in human cells Wang, Jianming Saponaro, Marco STAR Protoc Protocol G2/M DNA synthesis (G-MiDS) can be observed in one in five G2/M cells in unperturbed conditions by immunofluorescence microscopy. However, little is known of the genomic sites undergoing G-MiDS. Here, we describe a protocol which allows enriching for G2/M cells and investigating the sites of G-MiDS using BrdU-seq. This method can also be used to study the role of DNA replication or transcription-associated factors in affecting G-MiDS levels in different cell lines. For complete details on the use and execution of this protocol, please refer to Wang et al. (2021). Elsevier 2021-05-29 /pmc/articles/PMC8178111/ /pubmed/34136835 http://dx.doi.org/10.1016/j.xpro.2021.100570 Text en © 2021 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
Wang, Jianming
Saponaro, Marco
Protocol for analysis of G2/M DNA synthesis in human cells
title Protocol for analysis of G2/M DNA synthesis in human cells
title_full Protocol for analysis of G2/M DNA synthesis in human cells
title_fullStr Protocol for analysis of G2/M DNA synthesis in human cells
title_full_unstemmed Protocol for analysis of G2/M DNA synthesis in human cells
title_short Protocol for analysis of G2/M DNA synthesis in human cells
title_sort protocol for analysis of g2/m dna synthesis in human cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8178111/
https://www.ncbi.nlm.nih.gov/pubmed/34136835
http://dx.doi.org/10.1016/j.xpro.2021.100570
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