Illumina HiSeq 2500 sequencing of SSUB007616 : control_3
Briefly, 20 ng genomic DNA was subjected to MspI digestion (NEB, Beverly, MA, USA). End repair/dA-tailing reaction using Klenow Flagment (M0212S, NEB) and ligation by Illumina sequencing adapters using T4 DNA ligase (NEB). Then 150-350 bp fragments were gel-excised using NuSieve 3:1 agarose (Lonza Japan, Tokyo, Japan) and a MinElute Gel Extraction kit (QIAGEN, Valencia, CA, USA). The fragments were treated with sodium bisulfite using an EZ DNA Methylation-Gold Kit (Zymo Research, Orange, CA, USA). Library amplification and indexing were performed with KAPA HiFi HotStart Uracil+ ReadyMix (2x) (Kapa Biosystems, Woburn, MA).