UNIT 21.19 ChIP-Seq: A Method for Global Identification of Regulatory Elements in the Genome
Published Online: 1 JUL 2010
Copyright © 2010 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Raha, D., Hong, M. and Snyder, M. 2010. ChIP-Seq: A Method for Global Identification of Regulatory Elements in the Genome. Current Protocols in Molecular Biology. 91:21.19:21.19.1–21.19.14.
- Published Online: 1 JUL 2010
- Published Print: JUL 2010
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.
- mammalian cells;
- binding site;