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

2010; Nature Portfolio; Volume: 7; Issue: 2 Linguagem: Inglês

10.1038/nmeth.1414

1548-7105

Hongcang Gu, Christoph Bock, Tarjei S. Mikkelsen, Natalie Jäger, Zachary D. Smith, Eleni M. Tomazou, Andreas Gnirke, Eric S. Lander, Alexander Meissner,

Cancer-related gene regulation

Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. We optimized bisulfite sequencing for genome-scale analysis of clinical samples: here we outline how restriction digestion targets bisulfite sequencing to hotspots of epigenetic regulation and describe a statistical method for assessing significance of altered DNA methylation patterns. Thirty nanograms of DNA was sufficient for genome-scale analysis and our protocol worked well on formalin-fixed, paraffin-embedded samples.