title: Illumina HiSeq 2000 sequencing; Next generation sequencing identifies major DNA methylation changes during progression of Ph+ chronic myeloid leukemia_RRBS_samples
design: Next generation sequencing identifies major DNA methylation changes during progression of Ph+ chronic myeloid leukemia_RRBS_samples
library_construction_protocol: Mononuclear cells were isolated from peripheral blood or bone marrow using Ficoll. Genomic DNA (gDNA) was isolated from MNCs using QIAsymphony DSP DNA Midi Kit (Qiagen), MspI (New England Biolabs, NEB) digested, end-repaired and A-tailed using Klenow Polymerase (NEB) and dATP, dCTP and dGTP. Illumina adapters were ligated using Quick Ligase (NEB) followed by AMPure XP bead size selection (Beckman Coulter). RRBS libraries were bisulfite treated using EZ DNA MethylationDirect kit (Zymo Research) followed by quantification by qPCR. Enrichment PCR was performed using PfuTurbo Cx Hotstart kit (Agilent Technologies) followed by AMPure XP bead clean up. Quality of final libraries was checked by Experion analysis (BioRad).
attributes.tag: Experimental Factor: chronic myeloid leukemia
attributes.value: disease
attributes.tag: Experimental Factor: chronic phase chronic myeloid leukemia
attributes.value: disease staging