RRBS of human CLL sample: Sample CRC-0011-T-04
Peripheral blood mononuclear cells (PBMC) from normal donors and patients were isolated by Ficoll/Hypaque density gradient centrifugation. Mononuclear cells were used fresh or cryopreserved with 10% DMSO FBS and stored in vapor-phase liquid nitrogen until the time of analysis. RRBS libraries were generated from 25-100 ng of input DNA using the Ovation Methyl-Seq System (NuGen) following the manufacturer's recommendation. Libraries were sequenced per sample on an Illumina HiSeq2500. These reads were aligned to the human hg19 genome using BSmap with flags -v 0.05 -s 16 -w 100 -S 1 -p 8 -u. The methylation percentage for each CpG was calculated as the number of times the CpG appeared methylated divided by the total times the CpG was covered in sequencing reads.