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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| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725004137 |
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| Summary: | 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. |
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| ISSN: | 2666-1667 |