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An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions

Transcription factors are important regulators of cell fate and function. Knowledge about where transcription factors are bound in the genome is crucial for understanding their function. A common method to study protein-DNA interactions is chromatin immunoprecipitation (ChIP). Here, we present a rev...

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Detalles Bibliográficos
Autores principales: de Jonge, Wim J., Brok, Mariël, Kemmeren, Patrick, Holstege, Frank C.P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7357673/
https://www.ncbi.nlm.nih.gov/pubmed/32685929
http://dx.doi.org/10.1016/j.xpro.2020.100020
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author de Jonge, Wim J.
Brok, Mariël
Kemmeren, Patrick
Holstege, Frank C.P.
author_facet de Jonge, Wim J.
Brok, Mariël
Kemmeren, Patrick
Holstege, Frank C.P.
author_sort de Jonge, Wim J.
collection PubMed
description Transcription factors are important regulators of cell fate and function. Knowledge about where transcription factors are bound in the genome is crucial for understanding their function. A common method to study protein-DNA interactions is chromatin immunoprecipitation (ChIP). Here, we present a revised ChIP protocol to determine protein-DNA interactions for the yeast Saccharomyces cerevisiae. We optimized several aspects of the procedure, including cross-linking and quenching, cell lysis, and immunoprecipitation steps. This protocol facilitates sensitive and reproducible quantitation of protein-DNA interactions. For complete details on the use and execution of this protocol, please refer to (de Jonge et al., 2019).
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spelling pubmed-73576732020-07-17 An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions de Jonge, Wim J. Brok, Mariël Kemmeren, Patrick Holstege, Frank C.P. STAR Protoc Protocol Transcription factors are important regulators of cell fate and function. Knowledge about where transcription factors are bound in the genome is crucial for understanding their function. A common method to study protein-DNA interactions is chromatin immunoprecipitation (ChIP). Here, we present a revised ChIP protocol to determine protein-DNA interactions for the yeast Saccharomyces cerevisiae. We optimized several aspects of the procedure, including cross-linking and quenching, cell lysis, and immunoprecipitation steps. This protocol facilitates sensitive and reproducible quantitation of protein-DNA interactions. For complete details on the use and execution of this protocol, please refer to (de Jonge et al., 2019). Elsevier 2020-06-03 /pmc/articles/PMC7357673/ /pubmed/32685929 http://dx.doi.org/10.1016/j.xpro.2020.100020 Text en © 2020 The Author(s) http://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
de Jonge, Wim J.
Brok, Mariël
Kemmeren, Patrick
Holstege, Frank C.P.
An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions
title An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions
title_full An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions
title_fullStr An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions
title_full_unstemmed An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions
title_short An Optimized Chromatin Immunoprecipitation Protocol for Quantification of Protein-DNA Interactions
title_sort optimized chromatin immunoprecipitation protocol for quantification of protein-dna interactions
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7357673/
https://www.ncbi.nlm.nih.gov/pubmed/32685929
http://dx.doi.org/10.1016/j.xpro.2020.100020
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