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Analysis of the methylome of human embryonic stem cells employing methylated DNA immunoprecipitation coupled to next-generation sequencing

MPG-Autoren
http://pubman.mpdl.mpg.de/cone/persons/resource/persons50173

Grimm,  Christina
In vitro Ligand Screening (Zoltán Konthur), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons50054

Adjaye,  James
Molecular Embryology and Aging (James Adjaye), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Volltexte (frei zugänglich)

Grimm.pdf
(Verlagsversion), 416KB

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Zitation

Grimm, C., & Adjaye, J. (2012). Analysis of the methylome of human embryonic stem cells employing methylated DNA immunoprecipitation coupled to next-generation sequencing. In K. Turksen (Ed.), Methods in Molecular Biology (pp. 281-295). Humana Press.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-000E-F045-2
Zusammenfassung
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 (5 mC) 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).