title: GSM7024942: FB-Organoid D30 ETMR1 CP-673451; Homo sapiens; Mus musculus; RNA-Seq
library_construction_protocol: Cell dissociation using papain/DNAse solution, with incubation for 60min for organoids. Cell dissociation of 2D culture using Accutase and Trypsin EDTA for all conditions. Organoid samples had consistently > 90% vitality in manual counting, so they were not submitted to FACS sorting. 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. 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 the NextSeq2000 sequencing platform. scRNA-seq with Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1