Document Detail


PROTOCOLS: Chromatin Immunoprecipitation from Arabidopsis Tissues.
MedLine Citation:
PMID:  24653666     Owner:  NLM     Status:  PubMed-not-MEDLINE    
Abstract/OtherAbstract:
The ability of proteins to associate with genomic DNA in the context of chromatin is critical for many nuclear processes including transcription, replication, recombination, and DNA repair. Chromatin immunoprecipication (ChIP) is a practical and useful technique for characterizing protein / DNA association in vivo. The procedure generally includes six steps: (1) crosslinking the protein to the DNA; (2) isolating the chromatin; (3) chromatin fragmentation; (4) imunoprecipitation with antibodies against the protein of interest; (5) DNA recovery; and (6) PCR identification of factor associated DNA sequences. In this protocol, we describe guidelines, experimental setup, and conditions for ChIP in intact Arabidopsis tissues. This protocol has been used to study association of histone modifications, of chromatin remodeling ATPases, as well as of sequence-specific transcription factors with the genomic DNA in various Arabidopsis thaliana tissues. The protocol described focuses on ChIP-qPCR, but can readily be adapted for use in ChIP-chip or ChIP-seq experiments. The entire procedure can be completed within 3 days.
Authors:
Nobutoshi Yamaguchi; Cara M Winter; Miin-Feng Wu; Chang Seob Kwon; Dilusha A William; Doris Wagner
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Publication Detail:
Type:  Journal Article     Date:  2014-02-17
Journal Detail:
Title:  The Arabidopsis book / American Society of Plant Biologists     Volume:  12     ISSN:  1543-8120     ISO Abbreviation:  Arabidopsis Book     Publication Date:  2014  
Date Detail:
Created Date:  2014-03-21     Completed Date:  2014-03-21     Revised Date:  2014-03-24    
Medline Journal Info:
Nlm Unique ID:  101214894     Medline TA:  Arabidopsis Book     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e0170     Citation Subset:  -    
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