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Methylated DNA Immunoprecipitation (MeDIP) from Low Amounts of Cells

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Genomic Imprinting

Part of the book series: Methods in Molecular Biology ((MIMB,volume 925))

Abstract

Methylated DNA immunoprecipitation (MeDIP) is an immunocapturing approach for unbiased enrichment of DNA that is methylated on cytosines. The principle is that genomic DNA is randomly sheared by sonication and immunoprecipitated with an antibody that specifically recognizes 5-methylcytidine (5mC), which can be combined with PCR or high-throughput analysis (microarrays, deep sequencing). The MeDIP technique has been originally used to generate DNA methylation profiles on a genome scale in mammals and plants. Here we provide an optimized version of the MeDIP protocol suitable for low amounts of DNA, which can be used to study DNA methylation in cellular populations available in small quantities.

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References

  1. Meissner A (2010) Epigenetic modifications in pluripotent and differentiated cells. Nat Biotechnol 28:1079–1088

    Article  PubMed  CAS  Google Scholar 

  2. Laird PW (2010) Principles and challenges of genomewide DNA methylation analysis. Nat Rev Genet 11:191–203

    Article  PubMed  CAS  Google Scholar 

  3. Lister R, Pelizzola M, Dowen RH et al (2009) Human DNA methylomes at base resolution show widespread epigenomic differences. Nature 462:315–322

    Article  PubMed  CAS  Google Scholar 

  4. Weber M, Davies JJ, Wittig D et al (2005) Chromosome-wide and promoter-specific analyses identify sites of differential DNA methylation in normal and transformed human cells. Nat Genet 37:853–862

    Article  PubMed  CAS  Google Scholar 

  5. Down TA, Rakyan VK, Turner DJ et al (2008) A Bayesian deconvolution strategy for immunoprecipitation-based DNA methylome analysis. Nat Biotechnol 26:779–785

    Article  PubMed  CAS  Google Scholar 

  6. Pelizzola M, Koga Y, Urban AE et al (2008) MEDME: an experimental and analytical methodology for the estimation of DNA methylation levels based on microarray derived MeDIP-enrichment. Genome Res 18:1652–1659

    Article  PubMed  CAS  Google Scholar 

  7. Chavez L, Jozefczuk J, Grimm C et al (2010) Computational analysis of genome-wide DNA methylation during the differentiation of human embryonic stem cells along the endodermal lineage. Genome Res 20:1441–1450

    Article  PubMed  CAS  Google Scholar 

  8. Mohn F, Weber M, Schubeler D et al (2009) Methylated DNA immunoprecipitation (MeDIP). Methods Mol Biol 507:55–64

    Article  PubMed  CAS  Google Scholar 

  9. Borgel J, Guibert S, Li Y et al (2010) Targets and dynamics of promoter DNA methylation during early mouse development. Nat Genet 42:1093–1100

    Article  PubMed  CAS  Google Scholar 

  10. Jia J, Pekowska A, Jaeger S et al (2010) Assessing the efficiency and significance of methylated DNA immunoprecipitation (MeDIP) assays in using in vitro methylated genomic DNA. BMC Res Notes 3:240

    Article  PubMed  Google Scholar 

  11. Li N, Ye M, Li Y et al (2010) Whole genome DNA methylation analysis based on high throughput sequencing technology. Methods 52:203–212

    Article  PubMed  Google Scholar 

  12. Harris RA, Wang T, Coarfa C et al (2010) Comparison of sequencing-based methods to profile DNA methylation and identification of monoallelic epigenetic modifications. Nat Biotechnol 28:1097–1105

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by the CEFIC Long Research Initiative (LRI-EMSG49-CNRS-08).

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Correspondence to Michael Weber .

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Borgel, J., Guibert, S., Weber, M. (2012). Methylated DNA Immunoprecipitation (MeDIP) from Low Amounts of Cells. In: Engel, N. (eds) Genomic Imprinting. Methods in Molecular Biology, vol 925. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-011-3_9

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  • DOI: https://doi.org/10.1007/978-1-62703-011-3_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-010-6

  • Online ISBN: 978-1-62703-011-3

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