GSM4145774: Healthy 2_R848; Homo sapiens; RNA-Seq
isolation protocol: B-cells isolated by density centrifugation followed by negative selection with a memory B-cell isolation kit (Miltenyi) and subsequently depletion with CD3 and CD56 microbeads (Miltenyi). Alternatively, following density centrifugation, peripheral blood mononuclear cells (PBMCs) or bone marrow MNCs were initially depleted of cells expressing CD3 or CD56 with microbeads and then B-cells were positively selected with CD20 microbeads. RNA was extracted with TRIzol (Invitrogen) then subjected to DNAseI treatment (DNAfree, Ambion). RNA samples were quantified with a Qubit HS RNA assay kit (Thermo Fisher Scientific) and the quality was assessed by RNA HS assay using the TapeStation system (Agilent). Sequencing libraries were generated by using a TruSeq Stranded Total RNA Human/Mouse/Rat kit (Illumina) according to the manufacturer's protocol. Library performance was checked on the TapeStation followed by picogreen quantification. Equimolar pooling was performed across all the libraries and the final pool was sequenced on a NextSeq 500 platform (Illumina) using 76-bp single-end sequencing.
GEO Accession
GSM4145774