An optimized protocol for rapid, sensitive and robust on-bead ChIP-seq from primary cells

Integrative analysis of next-generation sequencing data can help understand dis-ease mechanisms. Specifically, ChIP-seq can illuminate where transcription regu-lators bind to regulate transcription. A major obstacle to performing this assay on primary cells is the low numbers obtained from tissues. The extensively vali-dated ChIP-seq protocol presented here uses small volumes and single-pot on -bead library preparation to generate diverse high-quality ChIP-seq data. This protocol allows for medium-to-high-throughput ChIP-seq of low-abundance cells and can also be applied to other mammalian cells. For complete details on the use and execution of this protocol, please refer to Brigidi et al. (2019), Carlin et al. (2018), Heinz et al. (2018), Nott et al. (2019), Sakai et al. (2019), and Seidman et al. (2020).

Automated summary

Integrative analysis of next-generation sequencing data is crucial for understanding disease mechanisms. ChIP-seq technique can reveal the binding sites of transcription regulators for transcriptional regulation. The validated ChIP-seq protocol presented here allows for medium-to-high-throughput analysis of low-abundance cells and can be applied to various mammalian cells.