Abstract
Eukaryotic transcriptional regulation is a tightly controlled process mediated by the binding of transcription factors (TFs) to the DNA as well as histone modifications. Chromatin immunoprecipitation and sequencing (ChIP-seq) is a technique to identify regions of DNA bound by TFs or marked by histone modifications and histone variants, providing essential insights into gene regulatory mechanisms. This chapter describes how to design a rigorous ChIP-seq experiment based on the cell or tissue type, the protein target, and the amount of material available. It also provides guidelines for library construction and sequencing. In addition, this chapter highlights best practices and common pitfalls in ChIP-seq data analysis, including quality control, peak calling, visualization, and differential analysis. Overall, this chapter outlines the key experimental steps and bioinformatics methods to obtain high-quality ChIP-seq data and analysis.
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