title: GSM8397945: PBMC_scRNAseq_LL_3; Homo sapiens; RNA-Seq
library_construction_protocol: fasting blood was collected into Vacutainer tubes containing anticoagulant citrate dextrose solution A (BD, 364606). Briefly, whole blood was processed within 2h of receipt and centrifuged at 480 g for 10 min. at room temperature (RT). Plasma was collected and the cell fraction diluted 1:2 in PBS, then placed in Ficoll-Paque PLUS density gradient media (GE Healthcare Life Sciences, 17144003) and centrifuged at 1,250 g for 20 min. at RT. The PBMC layer was collected, washed twice in PBS and resuspended at 2–5×105 cells per ml in 50% vol/vol RPMI-1640 (Gibco, 11875093) 40% vol/vol FBS 10% dimethyl sulfoxide (Tocris Bioscience, 3176) for frozen storage at –80 °C or liquid N2. On experiment date, specimens were thawed, washed and their viability assessed using a Cellometer Auto 2000 (Nexcellom). scRNA-seq libraries were prepared using the Chromium Single Cell 3' Library v3 reagent, Gel Bead and Multiplex Kit and Chip Kit (10X Genomics; 1000268, 1000215 and 100127, respectively) according to the manufacturer's instructions.