Protocol to apply spike-in ChIP-seq to capture massive histone acetylation in human cells

Di Wu; Liguo Wang; Haojie Huang

doi:10.1016/j.xpro.2021.100681

Protocol to apply spike-in ChIP-seq to capture massive histone acetylation in human cells

Research output: Contribution to journal › Article › peer-review

Abstract

Inhibition of histone deacetylases causes rapid and robust acetylation of histones. In this case, histone acetylation is likely increased on nearly every nucleosome, and the per-cell DNA/chromatin yield in chromatin immunoprecipitation (ChIP) experiments is significantly increased. Spike-in controls are essential for normalizing ChIP sequencing (ChIP-seq) data to capture this massive effect. Here, we report a detailed protocol of H3K27-ac ChIP-seq in human cells with chromatin from an ancestral species as a spike-in control. For complete details on the use and execution of this protocol, please refer to Wu et al. (2021).

Original language	English (US)
Article number	100681
Journal	STAR Protocols
Volume	2
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2021.100681
State	Published - Sep 17 2021

Keywords

Bioinformatics
Cell Biology
Cell-based Assays
ChIP-seq
Genomics
Sequence analysis

ASJC Scopus subject areas

General Immunology and Microbiology
General Biochemistry, Genetics and Molecular Biology
General Neuroscience

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10.1016/j.xpro.2021.100681

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abstract = "Inhibition of histone deacetylases causes rapid and robust acetylation of histones. In this case, histone acetylation is likely increased on nearly every nucleosome, and the per-cell DNA/chromatin yield in chromatin immunoprecipitation (ChIP) experiments is significantly increased. Spike-in controls are essential for normalizing ChIP sequencing (ChIP-seq) data to capture this massive effect. Here, we report a detailed protocol of H3K27-ac ChIP-seq in human cells with chromatin from an ancestral species as a spike-in control. For complete details on the use and execution of this protocol, please refer to Wu et al. (2021).",

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KW - Genomics

KW - Sequence analysis

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Protocol to apply spike-in ChIP-seq to capture massive histone acetylation in human cells

Abstract

Keywords

ASJC Scopus subject areas

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