Abstract
Interaction and co-occurrence of protein and DNA-based epigenetic modifications have become a topic of interest for many fundamental and biomedical questions. We describe within this chapter a protocol that combines two techniques in order to determine the methylation status of the DNA specifically associated with a protein of interest. First, DNA that directly interacts with the selected protein (such as a specific histone modification, a transcription factor, or any other DNA-associated protein) is purified by standard chromatin immunoprecipitation (ChIP). Second, the level of DNA methylation of this immunoprecipitated DNA is measured by bisulfite conversion and Pyrosequencing, a quantitative sequencing-by-synthesis method. This procedure allows determining the methylation status of genomic DNA associated to a specific protein at single nucleotide resolution.
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Acknowledgments
This work was supported by ATIP CNRS and Equipe d’Excellence Région Midi-Pyrenées grant to PBA. CM was recipient of an MNERT fellowship.
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Moison, C., Assemat, F., Daunay, A., Arimondo, P.B., Tost, J. (2015). DNA Methylation Analysis of ChIP Products at Single Nucleotide Resolution by Pyrosequencing® . In: Lehmann, U., Tost, J. (eds) Pyrosequencing. Methods in Molecular Biology, vol 1315. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2715-9_22
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DOI: https://doi.org/10.1007/978-1-4939-2715-9_22
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2714-2
Online ISBN: 978-1-4939-2715-9
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