GSM7814109: AD+no.11; Homo sapiens; Bisulfite-Seq
Genomic DNA (gDNA) was extracted from peripheral blood by using a Maxwell RSC Instrument (Promega, USA) with Maxwell RSC Buffy Coat DNA Kit (Promega, USA). Comprehensive methylation analysis of the 1st cohort was conducted by the target enrichment system, TruSeq methyl capture EPIC Kit (Illumina, Inc., San Diego, CA, USA), which was followed by next generation sequencing as follows: gDNA was fragmented to a few hundred base pairs (bps) by Covaris M220, followed by blunting, phosphorylation, addition of 3'-dA, and ligation of indexed adaptors to both ends of DNA.  DNA was mixed and hybridized to capture oligos, and hybridized DNA was recovered by streptavidin-conjugated magnetic beads.  Eluted DNA was treated with sodium bisulfite and amplified by PCR to construct a sequencing library, which was sequenced by using the Illumina HiSeq 2500 platform with paired-end reads of 100 bp by following the manufacturer's instructions.  Library construction and sequencing were conducted by Takara bio, inc. (Shiga, Japan).