Title

Chromatin immunoprecipitation (ChIP) coupled to detection by quantitative real-time PCR to study transcription factor binding to DNA in Caenorhabditis elegans

PubMed ID

18388953

UMMS Affiliation

Program in Gene Function and Expression

Date

4-5-2008

Document Type

Article

Subjects

Animals; Caenorhabditis elegans; Chromatin Immunoprecipitation; DNA; Gene Expression Regulation; Polymerase Chain Reaction; Protein Binding; Transcription Factors

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

In order to determine how signaling pathways differentially regulate gene expression, it is necessary to identify the interactions between transcription factors (TFs) and their cognate cis-regulatory DNA elements. Here, we have outlined a chromatin immunoprecipitation (ChIP) protocol for use in whole Caenorhabditis elegans extracts. We discuss optimization of the procedure, including growth and harvesting of the worms, formaldehyde fixation, TF immunoprecipitation and analysis of bound sequences through real-time PCR. It takes approximately 10-12 d to obtain the worm culture for ChIP; the ChIP procedure is spaced out over a period of 2.5 d with two overnight incubations.

Rights and Permissions

Citation: Nat Protoc. 2008;3(4):698-709. Link to article on publisher's site

Related Resources

Link to Article in PubMed