title: GSM5230717: HSPC_WT_UT_rep1; Mus musculus; RNA-Seq
library_construction_protocol: Briefly, bone marrow was flushed in PBS supplemented with 0.1% BSA and 2mM EDTA, filtered through 40μm cell-strainers, and subjected to red blood cell lysis in ACK buffer (0.15M NH4Cl, 10mM KHCO3, 0.1mM EDTA). The samples were stained with biotin anti-mouse Lineage Panel (BioLegend), PE-Cy7 cKit (clone 2B8, BioLegend), APC-Cy7 Sca-1 (D7, BioLegend), FITC CD48 (HM48-1, eBioscience), APC CD150 (TC15-12F12.2, BioLegend), PE Flt3 (A2F10, eBioscience), Brilliant Violet 421 CD34 (RAM34, BD Biosciences), and streptavidin-PECy5 (BioLegend). Cell sorting was performed on FACSAria, with FACS Diva software (BD Biosciences). The Hematopoietic stem and progenitor cells (HSPC) were sorted using FACS, gating on Lin-cKit+Sca1+CD150+. RNA was isolated using the MagMAX total RNA kit (Ambion, Life Technologies), and quality assessed using Bioanalyzer RNA Pico chips (Agilent). rRNA depletion and library preparation were performed using the SMARTer Stranded RNA-Seq kit (Takara Clontech). The libraries were sequenced on Illumina HiSeq 2000 and 2500 sequencers in paired-end 50bp configuration (two sequencing runs).