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. 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.
Gu
Hongcang
H
Broad Institute, Cambridge, Massachusetts, USA.
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
D001483:Base Sequence; D002874:Chromosome Mapping / Q000379:methods*; D003110:Colonic Neoplasms / Q000175:diagnosis* / Q000235:genetics*; D019175:DNA Methylation / Q000235:genetics*; D004273:DNA, Neoplasm / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004273:DNA, Neoplasm; D009711:Nucleotides