ChIP‐Seq: A Method for Global Identification of Regulatory Elements in the Genome
Open Access
- 15 July 2010
- journal article
- unit
- Published by Wiley in Current Protocols in Molecular Biology
- Vol. 91 (1) , 21.19.1-21.19.14
- https://doi.org/10.1002/0471142727.mb2119s91
Abstract
This unit describes ChIP‐Seq methodology, which involves chromatin immunoprecipitation (ChIP) followed by high‐throughput sequencing (Seq), and enables the genome‐wide identification of binding sites of transcription factors (TFs) and other DNA‐binding proteins. The process is initiated by cross‐linking DNA and DNA‐bound proteins. Subsequently, chromatin is isolated from nuclei and subjected to sonication. An antibody against a specific TF or DNA‐binding protein is then used to immunoprecipitate specific DNA‐TF complexes. ChIP DNA is purified, sequencing adapters are ligated, and 30‐ to 35‐nucleotide (nt) sequence reads are generated. The sequence of the DNA fragments is mapped back to the reference genome for determination of the binding sites. Curr. Protoc. Mol. Biol. 91:21.19.1‐21.19.14. © 2010 by John Wiley & Sons, Inc.Keywords
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