GSM8311015: Forebrain-Organoid D64; Homo sapiens; RNA-Seq
Cell dissociation using papain/DNAse solution, with incubation for 60min for organoids. Cell dissociation of 2D culture using Accutase. Approximately 10,000 vital single-cells of each sample were processed for single-cell RNA sequencing using the Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1, the Chromium Next GEM Chip G and the Single Index Kit T Set A (10X Genomics) according to the manufacturer's instructions. In brief, single-cell GEM (Gel Beads-in-Emulsion) were generated by the Chromium Controller, followed by GEM-RT (reverse transcription), Dynabeads cleanup, cDNA amplification, SPRIselect beads cleanup and library construction (for sample with a KK21 at the beginning we used single-indexed for all other samples, KK22-KK24 at the beginning, double-indexed libraries). Quality, purity, size and concentrations of cDNA and libraries were assured by measurement on a Tapestation 2200 (Agilent Technologies). Indexed libraries were sequenced separately by either the Illumina NextSeq 500 platform or the NextSeq2000 sequencing platform (please refer to "instrument model" in the sample description section). scRNA-seq with Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1