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

ABSTRACT: Integrative analysis of next-generation sequencing data can help understand disease mechanisms. Specifically, ChIP-seq can illuminate where transcription regulators bind to regulate transcription. A major obstacle to performing this assay on primary cells is the low numbers obtained from tissues. The extensively validated 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).

SUBMITTER: Texari L

PROVIDER: S-EPMC7921621 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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

Texari Lorane L Spann Nathanael J NJ Troutman Ty D TD Sakai Mashito M Seidman Jason S JS Heinz Sven S

STAR protocols 20210224 1

Integrative analysis of next-generation sequencing data can help understand disease mechanisms. Specifically, ChIP-seq can illuminate where transcription regulators bind to regulate transcription. A major obstacle to performing this assay on primary cells is the low numbers obtained from tissues. The extensively validated 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-t ...[more]

PMID: 33718886

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