An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions

STAR Protoc. 2022 Dec 16;3(4):101918. doi: 10.1016/j.xpro.2022.101918. Epub 2022 Dec 9.

Abstract

Genome-wide mapping of transcription factors (TFs) is critical to understand their functions. In chromatin immunoprecipitation (ChIP)-seq assay, it's challenging to study recruitment of low-abundant TFs transiently boud to the genome. Here, we present an optimized protocol using ChIP Next-Gen Seq Sepharose (Staph-seq) to efficiently pull down chromatin complexes. The double size selection promotes sensitive capture of genome-wide protein-DNA associations while eliminating potential Staph A contamination, which is a common problem in protocols using Staph A cells. For complete details on the use and execution of this protocol, please refer to Tao et al. (2020).1.

Keywords: Chromatin immunoprecipitation (ChIP); High Throughput Screening; Molecular Biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / genetics
  • Chromatin Immunoprecipitation / methods
  • DNA* / genetics
  • Sepharose
  • Transcription Factors* / genetics

Substances

  • Sepharose
  • DNA
  • Transcription Factors
  • Chromatin