Instrument: Illumina HiSeq 2500
Strategy: Bisulfite-Seq
Source: GENOMIC
Selection: RANDOM
Layout: SINGLE
Construction protocol: Qiagen Tissue DNA kit was used to isolate DNA from the tissue samples as per the manufacturer's protocol As per the protocol provided by Chatterjee A et al (PMID: 23193365). Briefly, 2.5µg of DNA was digested with 80U of MspI followed by end repair and A-tailing with clean up step. After ligation to methylated adapters, small fragments of DNA were removed by AMPure XP beads. Fragments of size 160-340bps (i.e. 40-220bps DNA + 120bp adapters) were extracted from 3% Nusieve gel using Qiaquick gel extraction kit. DNA samples were then treated for bisulfite conversion and clean-up. Libraries were prepared by13 cycles of PCR followed by size selection (160-340bps) from gel and, finally, assessed for quality using a Bioanalyzer and Kapa Kit.