Universität zu Köln

Rehimi, Rizwan, Bartusel, Michaela, Solinas, Francesca, Altmueller, Janine and Rada-Iglesias, Alvaro (2017). Chromatin Immunoprecipitation (ChIP) Protocol for Low-abundance Embryonic Samples. J. Vis. Exp. (126). CAMBRIDGE: JOURNAL OF VISUALIZED EXPERIMENTS. ISSN 1940-087X

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Abstract

Chromatin immunoprecipitation (ChIP) is a widely-used technique for mapping the localization of post-translationally modified histones, histone variants, transcription factors, or chromatin-modifying enzymes at a given locus or on a genome-wide scale. The combination of ChIP assays with next-generation sequencing (i.e., ChlP-Seq) is a powerful approach to globally uncover gene regulatory networks and to improve the functional annotation of genomes, especially of non-coding regulatory sequences. ChIP protocols normally require large amounts of cellular material, thus precluding the applicability of this method to investigating rare cell types or small tissue biopsies. In order to make the ChIP assay compatible with the amount of biological material that can typically be obtained in vivo during early vertebrate embryogenesis, we describe here a simplified ChIP protocol in which the number of steps required to complete the assay were reduced to minimize sample loss. This ChIP protocol has been successfully used to investigate different histone modifications in various embryonic chicken and adult mouse tissues using low to medium cell numbers (5 x 10(4) -5 x 10(5) cells). Importantly, this protocol is compatible with ChlP-seq technology using standard library preparation methods, thus providing global epigenomic maps in highly relevant embryonic tissues.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Rehimi, Rizwan UNSPECIFIED UNSPECIFIED UNSPECIFIED Bartusel, Michaela UNSPECIFIED UNSPECIFIED UNSPECIFIED Solinas, Francesca UNSPECIFIED UNSPECIFIED UNSPECIFIED Altmueller, Janine UNSPECIFIED UNSPECIFIED UNSPECIFIED Rada-Iglesias, Alvaro UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-224189
DOI:	10.3791/56186
Journal or Publication Title:	J. Vis. Exp.
Number:	126
Date:	2017
Publisher:	JOURNAL OF VISUALIZED EXPERIMENTS
Place of Publication:	CAMBRIDGE
ISSN:	1940-087X
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language TRANSCRIPTION FACTORS; CELL IDENTITY; ENHANCERS; H3K4ME3; SEQ; BINDING; GENOME; STATE Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/22418