Instrument: Illumina HiSeq 4000
Strategy: Bisulfite-Seq
Source: GENOMIC
Selection: RANDOM
Layout: PAIRED
Construction protocol: The DNA was extracted using Qiagen DNeasy Blood & Tissue Kits. After samples DNA testing, negative control DNAs were added into the initial DNA pool, and then fragmented into a 200-300bp range using Covaris S220. Next, terminal repairing, A-ligation, methylation sequencing adapters ligation were performed to DNA fragments. Then, DNA fragments undergone a bisulfite treatment using the EZ DNA Methylation Gold Kit from Zymo Research. Library concentration was firstly quantified by Qubit 2.0, and then is diluted to 1 ng/ul before an insert size verification using the Agilent Bioanalyzer 2100 and followed by Q-PCR. Finally, the library was paired-end sequenced using the Illumina HiSeq 4000.