title: GSM9191550: RC brain organoids, hiPSC line 1, day 90; Homo sapiens; RNA-Seq
library_construction_protocol: Organoids were cultured in Neurobasal A-Plus medium supplemented with B-27 Plus (2%) and Glutamax (1%) until harvesting at day 90. Pool of three organoids were separately incubated in Trypsin (Sigma Aldrich)/Accutase (StemCell Technologies) (1:1) containing 10 U/ml DNaseI (Thermo Fisher Scientific) in the gentleMACS™ Dissociator (Miltenyi Biotec) set at program NTDK1. After digestion the cell suspension was passed through a 70μm strainer. Samples were loaded to recover 16.000 cells onto a Chromium Next GEM Chip G Single Cell Kit (10x Genomics, PN-1000127) and processed through the Chromium controller to generate single-cell GEMs (Gel Beads in Emulsion). scRNA-seq libraries were prepared with the Chromium Next GEM Single Cell 3' Kit v3.1 (10x Genomics, PN-1000269). Organoids libraries were pooled and sequenced using the Element Biosciences AVITI. A total of 166 million reads were requested per library to ensure sufficient coverage for downstream analysis.
identifiers.namespace: GEO
identifiers.id: GSM9191550_r1