title: GSM6674754: Patient P1.1, mRNA-derived cDNA, scRNA-seq (capture reaction 1of2); Homo sapiens; RNA-Seq
library_construction_protocol: The frozen PBMCs were quickly thawed at 37°C and gently resuspended by serial additions of DMEM + 10% heat-inactivated FBS, for a final volume of 14ml. Following centrifugation at 300g for 5 min, cells were washed twice with 5ml DMEM + 10% HI-FBS to remove cell debris. Cell counting and viability assessment was done using the LIVE/DEAD™ Viability kit (ThermoFisher Scientific), according to manufacturer's guidelines. Single-cell library: The samples were loaded onto a 10X Genomics Chromium G chip aiming to capture 10,000 cells per samples. The reverse transcription and library preparation was performed using the Chromium Single Cell 3′ v2 Reagent Kits following manufacturers guidelines. The cDNA library quality was assessed using a TapeStation (Agilent). Sequencing: The cDNA libraries were sequenced on a HiSeq 4000 sequencer.