title: GSM8596782: PBMC, control, scRNAseq, subj2; Homo sapiens; RNA-Seq
library_construction_protocol: PBMCs were isolated from whole blood samples within 2 hours by Ficoll-Paque medium density gradient centrifugation for 30 min at 500 g. The cells were resuspended in 1X phosphate-buffered saline containing 0.04% filtered bovine serum albumin to achieve the required cell concentration (700–1200 cells per 1 μL). The final cell concentration and viability were determined using a Countess II Automated Cell Counter (Thermo Fisher Scientific, Waltham, MA, USA). Cell suspension with a concentration of 1000 cells per µl was used to achieve a targeted cell recovery up to 9000 cells. Gel beads-in-emulsion (GEMs) containing uniquely barcoded gel beads and suspended cells were generated using a 10×Genomics Chromium Controller device followed by reverse transcription. Single-cell libraries were constructed using Chromium Next GEM Single Cell 3ʹ Reagent Kit v3.1 according to the manufacturer's instruction. The main steps of library preparation included breaking GEMs, pooled cDNA fractions recovery, purification, and amplification, followed by enzymatic fragmentation, end-repair, A-tailing, adapter ligation, and final amplification. The quality of the single-cell libraries was assessed with Agilent 2200 TapeStation and Agilent High Sensitivity D1000 ScreenTape (both Agilent Technologies, Inc., Waltham, MA, USA).