GSM8499504: S3, scRNAseq; Homo sapiens; RNA-Seq
iPSC-derived NPCs or neurons were washed in PBS and detached as single cells by 5 minutes incubation with accutase. Single cells were suspended in PBS w/o Ca2+/Mg2+ containing 0.04% BSA, filtered using a 70 mm cell strainer Flowmi™ (734-2709, VWR), and counted with LUNA-II Automated Cell Counter (Logos Biosystems). Cell suspensions were loaded onto the Chromium Next GEM Chip G (10x Genomics) and run on the Chromium Controller (10x Genomics) to generate single-cell gel bead emulsion, according to the manufacturer's protocol (User Guide - Rev E). The Chromium Next GEM Single Cell 3' Kit v3.1 and the Chromium™ Next GEM Chip G Single Cell Kit (10x Genomics) were used to generate cDNA and the final libraries. The cDNA quality was assessed using high-sensitivity D5000 screen tape on TapeStation system 4150 (Agilent Technologies). Quality of libraries was assessed by using high sensitivity D1000 screen tape (Agilent Technologies). Finally, the libraries were sequenced on Novaseq6000 sequencer (Illumina) according to manufacturer specifications.