Abstract
Methylated DNA immunoprecipitation (MeDIP) is a versatile immunocapturing approach for unbiased detection of methylated DNA. In brief, genomic DNA is randomly sheared by sonication and immunoprecipitated with a monoclonal antibody that specifically recognizes 5-methylcytidine. The resulting enrichment of methylated DNA in the immunoprecipitated fraction can be determined by PCR to assess the methylation state of individual regions. Alternatively, MeDIP can be combined with large-scale analysis using microarrays as a genome-wide experimental readout. This protocol has been applied to generate comprehensive DNA methylation profiles on a genome-wide scale in mammals and plants, and further to identify abnormally methylated genes in cancer cells.
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Acknowledgements
The authors would like to thank the members of the Schübeler lab for helpful comments on the protocol.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Mohn, F., Weber, M., Schübeler, D., Roloff, TC. (2009). Methylated DNA Immunoprecipitation (MeDIP). In: Tost, J. (eds) DNA Methylation. Methods in Molecular Biology, vol 507. Humana Press. https://doi.org/10.1007/978-1-59745-522-0_5
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DOI: https://doi.org/10.1007/978-1-59745-522-0_5
Publisher Name: Humana Press
Print ISBN: 978-1-934115-61-9
Online ISBN: 978-1-59745-522-0
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