Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe
Summary: Chromatin immunoprecipitation (ChIP) assays provide quantitative information about the genomic localization of chromatin-binding proteins; however, their sensitivity is limited by several technical variables. Here, we present a protocol for developing and using Saccharomyces cerevisiae chro...
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| Language: | English |
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725004137 |
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| author | Jasbeer S. Khanduja Mo Motamedi |
| author_facet | Jasbeer S. Khanduja Mo Motamedi |
| author_sort | Jasbeer S. Khanduja |
| collection | DOAJ |
| description | Summary: Chromatin immunoprecipitation (ChIP) assays provide quantitative information about the genomic localization of chromatin-binding proteins; however, their sensitivity is limited by several technical variables. Here, we present a protocol for developing and using Saccharomyces cerevisiae chromatin as an exogenous spike-in control for ChIP of chromatin-binding proteins in Schizosaccharomyces pombe. We describe steps for preparing and validating the spike-in control. We then detail the procedure for adding spike-in control to S. pombe chromatin in ChIP assay and its use in data normalization.For complete details on the use and execution of this protocol, please refer to Khanduja et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-2b24e6e9806d4c7a84d96bf7eebdac21 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-2b24e6e9806d4c7a84d96bf7eebdac212025-08-20T03:58:21ZengElsevierSTAR Protocols2666-16672025-09-016310400710.1016/j.xpro.2025.104007Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombeJasbeer S. Khanduja0Mo Motamedi1Massachusetts General Brigham Krantz Family Center for Cancer Research and the Department of Medicine at Harvard Medical School, Charlestown, MA 02129, USAMassachusetts General Brigham Krantz Family Center for Cancer Research and the Department of Medicine at Harvard Medical School, Charlestown, MA 02129, USA; Corresponding authorSummary: Chromatin immunoprecipitation (ChIP) assays provide quantitative information about the genomic localization of chromatin-binding proteins; however, their sensitivity is limited by several technical variables. Here, we present a protocol for developing and using Saccharomyces cerevisiae chromatin as an exogenous spike-in control for ChIP of chromatin-binding proteins in Schizosaccharomyces pombe. We describe steps for preparing and validating the spike-in control. We then detail the procedure for adding spike-in control to S. pombe chromatin in ChIP assay and its use in data normalization.For complete details on the use and execution of this protocol, please refer to Khanduja et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725004137ChIP-seqMolecular BiologyChromatin immunoprecipitationChIP |
| spellingShingle | Jasbeer S. Khanduja Mo Motamedi Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe STAR Protocols ChIP-seq Molecular Biology Chromatin immunoprecipitation ChIP |
| title | Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe |
| title_full | Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe |
| title_fullStr | Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe |
| title_full_unstemmed | Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe |
| title_short | Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe |
| title_sort | protocol for development and use of spike in control for chromatin immunoprecipitation of chromatin binding proteins in schizosaccharomyces pombe |
| topic | ChIP-seq Molecular Biology Chromatin immunoprecipitation ChIP |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725004137 |
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