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ChIP-Seq: A Method for Global Identification of Regulatory Elements in the Genome.
Curr Protoc Mol Biol. 2010 Jul;Chapter 21:Unit21.19
Authors: Raha D, Hong M, Snyder M
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. (c) 2010 by John Wiley & Sons, Inc.
PMID: 20583098 [PubMed - in process]
More...
ChIP-Seq: A Method for Global Identification of Regulatory Elements in the Genome.
Curr Protoc Mol Biol. 2010 Jul;Chapter 21:Unit21.19
Authors: Raha D, Hong M, Snyder M
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. (c) 2010 by John Wiley & Sons, Inc.
PMID: 20583098 [PubMed - in process]
More...