Immunoprecipitation of Methylated DNA
DNA methylation contributes to the regulation of long-term gene repression by enabling the recruitment of transcriptional repressor complexes to methylated cytosines. Several methods for detecting DNA methylation at the gene-specific and genome-wide levels have been developed. Methylated DNA immunoprecipitation, or MeDIP, consists of the selective immunoprecipitation of methylated DNA fragments using antibodies to 5-methylcytosine. The genomic site of interest can be detected by PCR, hybridization to DNA arrays, or by direct sequencing. This chapter describes the MeDIP protocol and quality control tests that should be performed throughout the procedure.
Key wordsDNA methylation immunoprecipitation 5-methylcytosine antibody microarray
The basis for this MeDIP protocol has been the procedure established in Dirk Schübeler’s laboratory (Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland) by Michaël Weber and Dirk Schübeler and posted on the Epigenome Network of Excellence website (http://www.epigenome-noe.net/researchtools/protocol.php?protid=33). We are also grateful to Dirk Schübeler for discussion and advice. Our work is supported by the Research Council of Norway.
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