GSM5906147
The DNA was extracted using the PureLink Genomic DNA Mini Kit (Thermo, #K182002) according to manufacturer's instructions. Concentration of genomic DNA (gDNA) was quantified using a Qubit 3.0 Fluorometer. RRBS was performed on 100 ng gDNA of each sample using the NuGen Ovation® RRBS Methyl-Seq System (Tecan, #0353) following the manufacturer's recommendations with the following modifications (6). After the final repair step, the bisulfite conversion of DNA was conducted using the Qiagen EpiTect Fast Bisulfite Conversion kit (Qiagen, #59824) following the manufacturer's recommendations, eluting the bisulfite converted DNA in 23 µl EB. Libraries were amplified with 12 cycles of PCR. Amplified library purification with Agencourt RNAclean XP beads (Beckman Coulter, #A63987) was performed twice (1X). The purified libraries were quality-assessed on an Agilent 4150 Tape-Station HS D1000 ScreenTape and sequenced for 100-bp single-end reads on the NovaSeq6000 (Illumina).