title: GSM8210163: scRNA-seq_WT2_KEP2_GEX; Mus musculus; RNA-Seq
library_construction_protocol: Hematopoietic stem and progenitor cells (HSPCs) were enriched by neutrophil depletion. Bone marrow single cell suspensions were incubated with 10μg/ml anti-Ly6G (1A8, BioXcell) before neutrophil depletion using sheep anti-rat magnetic beads (Dynabeads, Life Technologies) and subsequently stained with cell surface markers. Cells were incubated with TotalSeq B anti-mouse hashtags (BioLegend) to enable de-multiplexing. Sorted single cell suspensions of CD34+cKIT+ cells were diluted to a final concentration of 1000 cells/μl in 1xPBS containing 0.04% BSA (400ug/ml). The Chromium X platform of 10X Genomics was used for single cell partitioning and barcoding. Per single cell suspension, each cell's transcriptome was barcoded during reverse transcription, pooled cDNA was amplified and Single Cell 3' Gene Expression libraries and Cell Hashing libraries via Feature barcode technology were prepared according to the manufacturer's protocol (CG000317, 10X Genomics). All libraries were quantified on a 2100 Bioanalyzer Instrument following the Agilent Technologies Protocol (Agilent DNA 7500 kit, G2938-90024). Sequence library pools were composed and quantified by qPCR, according to the KAPA Library Quantification Kit Illumina® Platforms protocol (KR0405, KAPA Biosystems).