Around 3×106 cells were harvested by 0.25 % trypsin digestion. DNA was extracted according to the instructions of Wizard® Genomic DNA Purification Kit (Promega A1125). Purity of DNA was determined by K5500 spectrophotometer, DNA quantification was performed using Qubit® 3.0 Fluorometer. DNA was digested by MspI followed by end-repairing. The 3’end of the end-repaired, phosphorylated fragments were tailed with dA, and the single A overhangs were ligated by the methylated adaptors. Library fragments of 40~220 bp were band isolated from 2% agarose gel, converted by bisulfite treatment, and amplified by PCR. The RRBS library was sequenced on an Illumina HiSeq2500 sequencers as paired-end 50 bp reads.
GEO Accession
GSM2443118