title: GSM4064464: N4; Homo sapiens; MeDIP-Seq
library_construction_protocol: Bone marrow buffy coats were collected from the CLL patients; the median lymphoid cell percentage was 85.75% (range, 41.60–99.00%). CD19 positive B-cells were collected from five healthy donors using magnetic bead sorting (EasySep; STEMCELL Technologies, Inc., Vancouver, Canada). Purity was confirmed using flow cytometry analysis (>95.0%). Genomic DNA was isolated using the Promega Maxwell 16 MDx Instrument. Genomic DNA (1 µg) was then sheared to 200-400 bp using a Covaris LE220 sonicator; the fragments were then subject to methyl-CpG enrichment using the Invitrogen MethylMiner Methylated DNA Enrichment Kit, which uses a recombinant form of human MBD protein-2. The enriched methylated DNA fragments were eluted as a single enriched population with a 2,000 mM NaCl elution buffer. The eluted DNA was then used to generate libraries according to the standard Illumina protocol. Briefly, the DNA fragments were subject to end repair, A-tailing of the 3' end, Illumina adapter ligation, size selection (aiming for 300-500 bp), PCR amplification, and validation using an Agilent Bioanalyzer. The libraries were then sequenced on the Illumina HiSeq 2000 platform.
identifiers.namespace: GEO
identifiers.id: GSM4064464