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20 February 2014 PROTOCOL: Chromatin Immunoprecipitation from Arabidopsis Tissues
Nobutoshi Yamaguchi, Cara M. Winter, Miin-Feng Wu, Chang Seob Kwon, Dilusha A. William, Doris Wagner
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Abstract

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.

© 2014 American Society of Plant Biologists
Nobutoshi Yamaguchi, Cara M. Winter, Miin-Feng Wu, Chang Seob Kwon, Dilusha A. William, and Doris Wagner "PROTOCOL: Chromatin Immunoprecipitation from Arabidopsis Tissues," The Arabidopsis Book 2014(12), (20 February 2014). https://doi.org/10.1199/tab.0170
Published: 20 February 2014
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