RNA-Seq of homo sapiens: Peripheral  Blood Mononuclear Cells
Aliquots of 10,000 cells were resuspended at 1000 cells per uL in sterile filtered 0.04% BSA in PBS. For higher cell yields (CD34- fraction), 35,000 cells were resuspended at 1000 cells per uL in 0.04% BSA in PBS. Single-cell libraries were prepared using 10X-3' v3.1 Gene Expression (10X Genomics) kit as per manufacturer's instruction. The libraries' quality and quantity were assessed using the TapeStation 4200 DNA High Sensitivity analysis and the Qubit 4 dsDNA high sensitivity assay, pooled and sequenced on the Illumina NovaSeq 6000 platform (Next Generation Sequencing Core, Salk Institute, La Jolla, CA, USA) using S4 flow cells (sequencing depth PE200) and 1% PhiX. For each sample, a custom calculation was done to assign a maximum number of reads based on the percentage of HSC present in the original sample resulting in range of 60 M to 500 M reads per sample.