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Book Sections Methods in Molecular Biology Year : 2015

DNA Methylation Analysis of ChIP Products at Single Nucleotide Resolution by Pyrosequencing®

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Abstract

Interaction and co-occurrence of protein and DNA-based epigenetic modifi cations 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 specifi cally associated with a protein of interest. First, DNA that directly interacts with the selected protein (such as a specifi c histone modifi cation, a transcription factor, or any other DNA-associated protein) is purifi ed by standard chromatin immunoprecipitation (ChIP). Second, the level of DNA methylation of this immunoprecipitated DNA is measured by bisulfi te conversion and Pyrosequencing, a quantitative sequencing-by-synthesis method. This procedure allows determining the methylation status of genomic DNA associated to a specifi c protein at single nucleotide resolution.
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Dates and versions

pasteur-03046253 , version 1 (08-12-2020)

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Céline Moison, Fanny Assemat, Antoine Daunay, Paola B Arimondo, Jörg Tost. DNA Methylation Analysis of ChIP Products at Single Nucleotide Resolution by Pyrosequencing®. Pyrosequencing, 1315, Springer, pp.315 - 333, 2015, Methods in Molecular Biology, 978-1-4939-2715-9. ⟨10.1007/978-1-4939-2715-9_22⟩. ⟨pasteur-03046253⟩
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