Abstract
The analysis of DNA-methylation on a genome-wide scale by next-generation sequencing techniques is an invaluable tool towards the understanding of the epigenetic basis of cellular differentiation. Methylated DNA immunoprecipitation (MeDIP) is an immunocapturing method using an antibody targeting 5-methylcytidine (5mC) and thereby enriching methylated DNA. MeDIP combined with next-generation sequencing (MeDIP-seq) provides a powerful tool for the analysis of genome-wide DNA-methylation profiles. Here, we describe a protocol for the preparation of MeDIP samples suitable for next-generation sequencing on a Genome Analyser (Illumina).
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Acknowledgments
CG and JA are funded by the German Ministry of Education and Research (BMBF) within its National Genome Research Network (01GS08111) and the ERASysBio Plus programme (0315717A). We thank Justyna Jozefczuk for providing genomic DNA of undifferentiated human embryonic stem cells and the corresponding endoderm-differentiated cells used for the qPCR assay in Fig. 3 and Aleksey Soldatov, Tatiana Borodina and Aleksey Kirpy for advice on library preparation. We thank Vardhman Rakyan and Stephan Beck for helpful discussions regarding the MeDIP-seq protocol and for providing the human control primer sequences prior to publication.
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Grimm, C., Adjaye, J. (2012). Analysis of the Methylome of Human Embryonic Stem Cells Employing Methylated DNA Immunoprecipitation Coupled to Next-Generation Sequencing. In: Turksen, K. (eds) Human Embryonic Stem Cells Handbook. Methods in Molecular Biology, vol 873. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-794-1_19
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DOI: https://doi.org/10.1007/978-1-61779-794-1_19
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