GSM6346363: Nup358 knock-out, replicate 2, scRNA-seq; Mus musculus; RNA-Seq
Bone marrow cells were stained for flow cytometry with Lin antibody cocktail and with antibodies against Sca1 and cKit. Before sorting, cells were resuspended in HBSS, 1% BSA, 20 mM HEPES pH 7.2. Live Lin-Sca1+cKit+ cells were sorted at 4°C on a BD FACSAria II Cell Sorter (BD Biosciences) with a 70-μm nozzle into DMEM without glucose, glutamine, sodium pyruvate, phenol red and supplemented with 10% FBS. Purity was verified after each sort and was found to be >99%. Sorted LSK cells were loaded into a Chromium Next GEM Chip and processed with the Chromium Controller (10x Genomics) according to the manufacturer's instructions for single cell barcoding. Libraries were prepared using Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10x Genomics) and were sequenced to an average depth of about 50,000 reads per cell.