title: GSM3099233: ID236 CMP; Homo sapiens; RNA-Seq
library_construction_protocol: After mobilization with granulocyte-colony stimulating factor (G-CSF), stem-cell enriched mononuclear leukocytes were obtained via plasmapheresis from peripheral blood of donors for clinical stem cell transplantation (Clinical Hematology, University Hospital Zurich, Zurich Switzerland). Left-over material after clinical transplantation was enrichted for CD34+ stem/progenitor cells using immunomagnetic beads according to manufacturer instructions (CD34 MicroBead Kit; Miltenyi Biotec). Fluorescence-activated cell sorting and antibodies specific for CD2, CD3, CD4, CD7, CD8, CD14, CD19, CD56, CD10, CD11b, CD20, CD235a, CD34, CD38, CD123, CD45RA were used to isolate the various cell types (HSC, CMP, GMP, MEP). 10'000 cells of each subpopulations were sorted into RNeasy lysis buffer containing beta-mercaptoethanol. Total RNA was purified according to manufacturer instructions using the RNeasy Plus Micro Kit (Qiagen). The samples were of high enough quality for a poly-A tail based approach for RNA sequencing. RNA sequencing was performed as described in Picelli S. et al. Nature Protocols 2014 using the Illumina HiSeq 4000 sequencing platform. RNA libraries were prepared for sequencing using standard Illumina protocols.
attributes.tag: GEO Accession
attributes.value: GSM3099233