Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution

Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. Here, we optimize bisulfite sequencing for genome-scale analysis of clinical samples. Specifically, we outline how restriction digestion targets bi...

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Detalles Bibliográficos
Autores principales: Gu, Hongcang, Bock, Christoph, Mikkelsen, Tarjei S., Jäger, Natalie, Smith, Zachary D., Tomazou, Eleni, Gnirke, Andreas, Lander, Eric S., Meissner, Alexander
Formato: Texto
Lenguaje:English
Publicado: 2010
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2860480/
https://www.ncbi.nlm.nih.gov/pubmed/20062050
http://dx.doi.org/10.1038/nmeth.1414
Descripción
Sumario:Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. Here, we optimize bisulfite sequencing for genome-scale analysis of clinical samples. Specifically, we outline how restriction digestion targets bisulfite sequencing to hotspots of epigenetic regulation; we show that 30ng of DNA are sufficient for genome-scale analysis; we demonstrate that our protocol works well on formalin-fixed, paraffin-embedded (FFPE) samples; and we describe a statistical method for assessing significance of altered DNA methylation patterns.

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