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ChIP-Seq Analysis of Histone Modifications at the Core of the Arabidopsis Circadian Clock

  • Jordi Malapeira
  • Paloma MasEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1158)

Abstract

Over the past years, chromatin modification has emerged as a key regulator of gene expression. A very useful method for chromatin analysis is chromatin immunoprecipitation (ChIP), which allows the quantification and localization of specific histone modifications. The basic steps of the ChIP protocol include cross-linking of histones and DNA, chromatin isolation, shearing the DNA into smaller fragments, immunoprecipitation with specific antibodies, and enrichment analysis by several methods including real-time quantitative PCR (ChIP-qPCR), microarray hybridization (ChIP-chip), or sequencing (ChIP-seq). Here, we describe how to use ChIP-qPCR to analyze histone modifications at the core of the Arabidopsis thaliana circadian clock. We also briefly discuss a number of protocol adjustments to be considered in ChIP-seq experiments.

Key words

Chromatin immunoprecipitation (ChIP) ChIP-qPCR Circadian clock Histone modifications Arabidopsis thaliana 

Notes

Acknowledgements

The work in our laboratory is supported by grants from the Ramón Areces Foundation, the Spanish Ministry of Science and Innovation (MICINN) (BIO2010-16483), the EMBO YIP program and from EUROHORCS (European Heads of Research Councils) and the European Science Foundation (ESF) through the EURYI Award to Paloma Mas. We thank Rossana Henriques for critical comments on the manuscript.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Center for Research in Agricultural Genomics (CRAG), Consortium CSIC-IRTA-UAB-UB, Parc de Recerca UAB, Edifici CRAG, Campus UAB, Bellaterra (Cerdanyola del Vallés)BarcelonaSpain

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