GSM2100959: IgM+ memory B cells replicate2; Mus musculus; RNA-Seq
Spleen cells were isolated and all mononuclear cells were stained with a combination of antibodies (anti-CD19, anti-IgM, anti-IgG). Then subsets of memory B cells were sorted as CD19+HEL+YFP+IgM+ or IgG+ and naive cells as CD19+HEL+YFP-IgM+ using a FACSAriaIII flow cytometer. The purity of the sorted populations varied from 90-95%. Directional libraries were prepared using the TruSeq Stranded total RNA Sample preparation kit following the manufacturer’s instructions (Illumina). The libraries were quantified by fluorimetric measurements with the Qubit® dsDNA HS Assay Kit (ThermoFisher). 51-bp Single Read sequences were generated on the Hiseq2500 sequencer according to manufacturer’s instructions (Illumina).  Sequences were demultiplexed using the Illumina pipeline (Gerald, included in CASAVA version 1.7) giving FASTQ formatted reads.
GEO
GSM2100959