title: GSM8241615: CDG-12105,scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: At day 92, 4 human corical brain organoids per cell line (CDG-0458, CDG-1017, CDG-11740, CTRL-8856.3, CTRL-1363.1, CTRL-8399) were dissociated into a single cell suspension using the Miltenyi Biotec Neurosphere Dissociation Kit (P) and resuspended  in 1x PBS + 0.04% BSA The cDNA master mix was prepared according to the manufacture's instruction for Chromium Single Cell 3' Library and Gel Bead Kit (10x Genomics). Per sample the aim was to recover 10,000 cells and as such the cDNA master mix was added to the cells so that we have 500,000 cell per ml.  Subsequently, these cells, Gel Beads, and partitioning oil were added to the Chromium Single Cell G chip. The filled chip was loaded into the Chromium X Controller, where each sample was processed and the individual cells within the sample were partitioned into uniquely labeled Gel Beads-In-Emulsion (GEMs).  The GEMs were collected from the chip and taken to the bench for reverse transcription, GEM dissolution, and cDNA clean-up. The resulting cDNA contained a pool of uniquely barcoded molecules, and a portion of this cDNA was taken for library construction. To create gene expression libraries, standard Illumina sequencing primers and unique sample index (Dual Index Kit TT-Set A; 10x Genomics) were added to each cDNA pool. All cDNA pools and resulting libraries were quantified using Qubit High Sensitivity assays (Thermo Fisher Scientific) and Agilent Bioanalyzer High Sensitivity chips (Agilent). The libraries were subsequently sequenced at 50,000 fragment reads per cell by using the Illumina's standard protocol and the Illumina NovaSeq™ 6000 S4 flow cell