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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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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). |
format | Online Article Text |
id | pubmed-7357673 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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