Chapter title |
DNA Methylation Analysis of ChIP Products at Single Nucleotide Resolution by Pyrosequencing(®).
|
---|---|
Chapter number | 22 |
Book title |
Pyrosequencing
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2715-9_22 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2714-2, 978-1-4939-2715-9
|
Authors |
Moison, Céline, Assemat, Fanny, Daunay, Antoine, Arimondo, Paola B, Tost, Jörg, Céline Moison, Fanny Assemat, Antoine Daunay, Paola B. Arimondo, Jörg Tost, Arimondo, Paola B. |
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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