Genomic DNA for DNA methylation profiling was isolated using the DNeasy Blood & Tissue Kit (Qiagen). DNA was fragmented by BioRuptor sonication (Diagenode, Denville, NJ). Fragmented DNA was analyzed with bioanalyzer, and 1microgram of sonicated DNA underwent methyl-CpG enrichment using the MethylMiner™ Methylated DNA Enrichment Kit (Invitrogen). Library preparation was undertaken using 5 ng of methyl-enriched DNA using the NEBNext DNA Library Preparation Kit for Illumina (New England Biolabs). Library DNA was quantified with MultiNA capillary electrophoresis instrument with DNA-500 kit. Libraries were sequenced at a concentration of 10 pM on Genome Analyzer IIx instrument for 36 cycles. Illumina base-calling was performed off-line with OLB v1.8.
GEO Accession
GSM1462880