GSM8389442: SCO organoid, Ctrl, scRNAseq; Homo sapiens; RNA-Seq
For each line, 14 organoids were pooled and minced into <1 mm pieces. These pieces were enzymatically digested with papain dissociation system (Worthington) and incubated with DNase for 40min. Then, the organoids were mechanically dissociated with 5mL pipette, filtered with 40uM mesh, then centrifiged and washed with DMEM twice. After cell counting, the cells were suspened in DMEM/10% FBS at a concentration of 1000 cells/uL. Library constructed with Chormium Next GEM Single Cell 3' Reagent Kits v 3.1. The cells were combined with RT reagents and loaded onto 10X Next GEM Chip with gel beads. ~100uL of emulsion was taken from the chip and incubated to generated barcoded cDNA from each cell. The samples were then cleaned with SPRI select beads.