title: GSM1412559: WT rep1_LSK; Mus musculus; RNA-Seq
library_construction_protocol: LSKs (FACS sorted from freshly isolated bone marrow mononuclear cells), naive splenic B cells (freshly FACS sorted) or FACS sorted LPS-activated B cells were used. Cells from individual animals (4 per genotype/ per experiment) were pooled, and total RNA was harvested using Trizol reagent. RNAseq was performed on an Illumina Hiseq2000 machine. Naive splenic B cells were obtained by CD43 depletion using biotynilated anti-CD43 (Clone S7, BD Biosciences), and the Imag system (BD Biosciences). Naive B cells were directly FACS sorted with fluorochrome-conjugated antibody specific for CD19, or cultured in vitro for four days in IMDM+8%FBS and 50 microgram/ml E.coli LPS (Sigma) to obtain activated B cells, followed by sorting. Two independent experiments were performed. Cell sorting was performed by FACSAria (BD Biosciences). RNA libraries were prepared for sequencing using standard Illumina TruSeq RNA Sample Prep protocols.
attributes.tag: GEO Accession
attributes.value: GSM1412559