GSM6256043: Tubuloids FAF+Li, scRNAseq; Homo sapiens; RNA-Seq
BME was removed by treatment with 1 mg/mL dispase II for 30 min. at 37 °C. Tubuloids were washed and dissociated using accutase at 37 °C and mechanical shearing using a flamepolished pipette. When single cells, tubuloids were washed, counted and resuspended in PBS without calcium and magnesium. -20 °C methanol was added dropwise to the samples to achieve a 80% methanol solution and then the samples were stored at -80 °C. Library preparation and sequencing were done by Single Cell Discoveries (https://www.scdiscoveries.com/). Library preparation was performed by SingleCellDiscoveries (10x Genomics). Briefly, a pool of Gel Beads coated with barcoded primers, cells and enzymes were loaded on the 10x Genomics microfluidics chip and placed in the chromium controller. Within the chromium controller, Gel Bead-in-emulsion (GEMs) formed. Gel beads were dissolved and molecules from a single cell were captured and barcoded. Barcoded fragments were pooled, and sequencing libraries were made. After sequencing, reads were mapped back to the corresponding single cell using STAR (v. 2.6.1)