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Methylation Profiling Using Methylated DNA Immunoprecipitation and Tiling Array Hybridization

  • Hoi-Hung Cheung
  • Tin-Lap Lee
  • Owen M. Rennert
  • Wai-Yee Chan
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 825)

Abstract

DNA methylation is an important epigenetic modification that regulates development and plays a role in the pathophysiology of many diseases. It is dynamically changed during germline development. Methylated DNA immunoprecipitation (MeDIP) is an efficient, cost-effective method for locus-specific and genome-wide analysis. Methylated DNA fragments are enriched by a 5-methylcytidine-recognizing antibody, therefore allowing the analysis of both CpG and non-CpG methylation. The enriched DNA fragments can be amplified and hybridized to tiling arrays covering CpG islands, promoters, or the entire genome. Comparison of different methylomes permits the discovery of differentially methylated regions that might be important in disease- or tissue-specific expression. Here, we describe an established MeDIP protocol and tiling array hybridization method for profiling methylation of testicular germ cells.

Key words

MeDIP DNA methylation Tiling arrays 

Notes

Acknowledgments

This work was supported in part by the Intramural Research Program of the National Institutes of Health (NIH), Eunice Kennedy Shriver National Institute of Child Health and Human Development, and in part by the Chinese University of Hong Kong.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Hoi-Hung Cheung
    • 1
  • Tin-Lap Lee
    • 2
    • 1
  • Owen M. Rennert
    • 1
  • Wai-Yee Chan
    • 3
  1. 1.Section on Clinical and Developmental GenomicsEunice Kennedy Shriver National Institute of Child Health and Human DevelopmentBethesdaUSA
  2. 2.School of Biomedical SciencesThe Chinese University of Hong KongHong KongChina
  3. 3.School of Biomedical SciencesThe Chinese University of Hong KongHong Kong SARChina

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