Protocol for development and use of spike-in control for chromatin immunoprecipitation of chromatin-binding proteins in Schizosaccharomyces pombe.

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).