GSM9830405: 20086-01-01; Homo sapiens; RNA-Seq
Peripheral blood mononuclear cells (PBMCs) were isolated from EDTA whole blood on Histopaque-1077. After washing, cells were counted, and 3-5 million cells were resuspended in sterile freezing media (90% FBS and 10% DMSO). Cells were slowly frozen in an isopropanol-containing cryopreservation vessel at -80C for 24 h then stored in liquid nitrogen. Final library was constructed using 10X Genomics 3' GEX v3.1 (10x Genomics) based on manufacturer's manual. The quality of FL was assessed by High Sensitivity D5000 Tapestation (Agilent Technologies Inc., California, USA) and quantified by Qubit 2.0 DNA HS assay (ThermoFisher, Massachusetts, USA). The average final library size is 400bp with an insert size of about 250bp. Equimolar pooling of libraries was performed based on QC values and sequenced on an Illumina® Hiseq X (Illumina, California, USA) with a read length configuration of 150 PE targeting for 100M total reads per sample (50M in each direction).