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An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions
Genome-wide mapping of transcription factors (TFs) is critical to understand their functions. In chromatin immunoprecipitation (ChIP)-seq assay, it’s challenging to study recruitment of low-abundant TFs transiently boud to the genome. Here, we present an optimized protocol using ChIP Next-Gen Seq Se...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9763742/ https://www.ncbi.nlm.nih.gov/pubmed/36595937 http://dx.doi.org/10.1016/j.xpro.2022.101918 |
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author | Tao, Fang Rhonda, Egidy He, Xi Perry, John M. Li, Linheng |
author_facet | Tao, Fang Rhonda, Egidy He, Xi Perry, John M. Li, Linheng |
author_sort | Tao, Fang |
collection | PubMed |
description | Genome-wide mapping of transcription factors (TFs) is critical to understand their functions. In chromatin immunoprecipitation (ChIP)-seq assay, it’s challenging to study recruitment of low-abundant TFs transiently boud to the genome. Here, we present an optimized protocol using ChIP Next-Gen Seq Sepharose (Staph-seq) to efficiently pull down chromatin complexes. The double size selection promotes sensitive capture of genome-wide protein-DNA associations while eliminating potential Staph A contamination, which is a common problem in protocols using Staph A cells. For complete details on the use and execution of this protocol, please refer to Tao et al. (2020).(1) |
format | Online Article Text |
id | pubmed-9763742 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-97637422022-12-21 An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions Tao, Fang Rhonda, Egidy He, Xi Perry, John M. Li, Linheng STAR Protoc Protocol Genome-wide mapping of transcription factors (TFs) is critical to understand their functions. In chromatin immunoprecipitation (ChIP)-seq assay, it’s challenging to study recruitment of low-abundant TFs transiently boud to the genome. Here, we present an optimized protocol using ChIP Next-Gen Seq Sepharose (Staph-seq) to efficiently pull down chromatin complexes. The double size selection promotes sensitive capture of genome-wide protein-DNA associations while eliminating potential Staph A contamination, which is a common problem in protocols using Staph A cells. For complete details on the use and execution of this protocol, please refer to Tao et al. (2020).(1) Elsevier 2022-12-09 /pmc/articles/PMC9763742/ /pubmed/36595937 http://dx.doi.org/10.1016/j.xpro.2022.101918 Text en © 2022 The Authors 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 Tao, Fang Rhonda, Egidy He, Xi Perry, John M. Li, Linheng An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions |
title | An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions |
title_full | An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions |
title_fullStr | An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions |
title_full_unstemmed | An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions |
title_short | An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions |
title_sort | optimized chromatin immunoprecipitation protocol using staph-seq for analyzing genome-wide protein-dna interactions |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9763742/ https://www.ncbi.nlm.nih.gov/pubmed/36595937 http://dx.doi.org/10.1016/j.xpro.2022.101918 |
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