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

ABSTRACT: 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.

SUBMITTER: Gu H 

PROVIDER: S-EPMC2860480 | biostudies-literature | 2010 Feb

REPOSITORIES: biostudies-literature

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

Gu Hongcang H   Bock Christoph C   Mikkelsen Tarjei S TS   Jäger Natalie N   Smith Zachary D ZD   Tomazou Eleni E   Gnirke Andreas A   Lander Eric S ES   Meissner Alexander A  

Nature methods 20100110 2

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 anal  ...[more]

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