GSM4118494: CD14_E_g_32; Homo sapiens; RNA-Seq
Peripheral blood mononuclear cells (PBMC) were isolated using Ficoll-Paque™ (GE healthcare life Sciences, Little Chalfont, United Kingdom). The PBMCs were subsequently sorted using Magnetic Cell Isolation and Cell Separation (AutoMACS Pro 360; Miltenyi Biotec, Bergisch Gladbach, Germany), positively selected for monocytes using CD14 MicroBeads, human, and negatively selected for CD4+ T cells using the CD4+ T Cell Isolation Kit, human (both from Miltenyi Biotec), according to the manufacturer's protocol. RNA was extracted using the RNeasy Mini Kit (Qiagen) and tested for quantity and purity by Nanodrop (Thermo Scientific) and quality by RIN-value with Agilent 2100 Bioanalyzer (Agilent technologies, Inc., Santa Clara, Ca, USA). All Samples were aliquoted and stored per 1.5-2.0 µg at -80°C until use. Total RNA was sent to GenomeScan B.V. (Leiden, Netherlands) for library preparation and sequencing. Before sequencing, the RNA sample preparation was performed using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England BioLabs) according to manufacturer's protocol. Briefly, mRNA was isolated from total RNA using the oligo-dT magnetic beads. After fragmentation of the mRNA, a cDNA synthesis was performed.
GEO Accession
GSM4118494