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Elife. 2015 Jun 16;4:e09225. doi: 10.7554/eLife.09225.

A simple method for generating high-resolution maps of genome-wide protein binding.

Author information

1
Fred Hutchinson Cancer Research Center, Seattle, United States.

Abstract

Chromatin immunoprecipitation (ChIP) and its derivatives are the main techniques used to determine transcription factor binding sites. However, conventional ChIP with sequencing (ChIP-seq) has problems with poor resolution, and newer techniques require significant experimental alterations and complex bioinformatics. Previously, we have used a new crosslinking ChIP-seq protocol (X-ChIP-seq) to perform high-resolution mapping of RNA Polymerase II (Skene et al., 2014). Here, we build upon this work and compare X-ChIP-seq to existing methodologies. By using micrococcal nuclease, which has both endo- and exo-nuclease activity, to fragment the chromatin and thereby generate precise protein-DNA footprints, high-resolution X-ChIP-seq achieves single base-pair resolution of transcription factor binding. A significant advantage of this protocol is the minimal alteration to the conventional ChIP-seq workflow and simple bioinformatic processing.

KEYWORDS:

CTCF; D. melanogaster; RNA polymerase II; chromosomes; evolutionary biology; genes; genomics; human; transcription factors

PMID:
26079792
PMCID:
PMC4480131
DOI:
10.7554/eLife.09225
[Indexed for MEDLINE]
Free PMC Article

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