alias: GSM1564141
title: GSM1564141: CD34_iPSC_RRBS_2; Homo sapiens; Bisulfite-Seq
library_construction_protocol: Murine hematopoietic stem cells (LSK) were isolated using cell sorting (FACS ARIA II, BD) using fluorochrome-conjugated antibodies against Sca1 and cKit. After sorting samples were immeditaley processed for methylation analysis DNA was digested with MspI (NEB), a methylation-insensitive enzyme that cuts C’CGG. Digested DNA was size selected on a 4% NuSieve 3:1 Agarose gel (Lonza). For each sample, two slices containing DNA fragments of 40-120 bp and 120-220 bp, respectively, were excised from the unstained preparative portion of the gel. These two size fractions were kept apart throughout the procedure including the final sequencing. Pre-annealed Illumina adaptors containing 5’-methyl-cytosine instead of cytosine were ligated to size-selected MspI fragments. Adapter-ligated fragments were bisulfite-treated using the EZ DNA Methylation kit (Zymo Research, Orange, CA). The products were PCR amplified, size selected before subjecting to sequencing.
identifiers.namespace: GEO
identifiers.id: GSM1564141
attributes.tag: GEO Accession
attributes.value: GSM1564141