- An Optimized Protocol for ChIP-Seq from Human Embryonic Stem Cell Cultures.
An Optimized Protocol for ChIP-Seq from Human Embryonic Stem Cell Cultures.
STAR protocols (2020-10-02)
Adrienne E Sullivan, Silvia D M Santos
PMID33000002
ABSTRACT
Chromatin immunoprecipitation (ChIP) followed by next-generation sequencing is a powerful technique that characterizes the genome-wide DNA-binding profile of a protein of interest. The general ChIP-seq workflow has been applied widely to many sample types and target proteins, but sample-specific optimization of various steps is necessary to achieve high-quality data. This protocol is specifically optimized for cultured human embryonic stem cells (hESCs), including steps to check sample quality and non-specific enrichment of "hyper-ChIPable" regions prior to sequencing. For complete details on the use and execution of this protocol, please refer to Gunne-Braden et al. (2020).
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Sodium chloride, Molecular Biology, DNase, RNase, and protease, none detected, ≥99% (titration)