title: GSM4725195: Bulk_SERIES_1: MPP_BCG_1; Mus musculus; RNA-Seq
library_construction_protocol: Bone marrow cells were harvested from femurs, tibiae, humeri and ilia, and incubated with the described biotin-conjugated antibodies against lineage-committed cells. Subsequently, the cells were incubated with MACS streptavidin conjugated microbeads (Miltenyi) for 30 minutes at 4°C. A depletion of the lineage-committed cells was performed using AutoMACS (Miltenyi) to reduce the contaminating total cell number. The remaining cells were incubated with the corresponding antibodies for HSCs vs MPPs cells. Total RNA was extracted from the sorted HSCs and MPPs using Magmax-96 Total RNA isolation kit (life technologies, Ambion 1830M). RNA quality was evaluated spectrophotometrically, and quality was assessed with the Agilent 2100 Bioanalyzer RNA-sequencing libraries were prepared using the SMARTer® Stranded RNA-­Seq Kit (for HSCs) and sequenced with single-­end 100bp reads on an Illumina HiSeq2500.