title: Ctrl1
organism: Homo sapiens
description: Protocols: For each of the 3 control and 3 homozygous mutant samples, 10 6 week-old organoids were pooled, while 2 organoids for used for each sample for the 10 week-old single cell analysis. The organoids were minced into small pieces using a sterile razor blade and dissociated into single cell suspensions using a Worthington Papain Dissociation kit (Worthington Biochemical, #LK003150) as per manufacturer's instruction, except for D94 organoids for which the Papain incubation time was increased to 75 minutes. After papain treatment, cells were centrifuged, resuspended in ice-cold 1x PBS and filtered using 40mm pluriStrainer Mini filter (Fisher Scientific, #431004050) and the final cell concentration was adjusted to 1000 cells/ml in PBS. Single-cell RNA-Seq libraries were prepared for the 10x scRNA sequencing platform according to the manufacturer's instructions with the Chromium Next GEM Single Cell 3@ Reagent Kit v3.1 (Dual Index; 10x Genomics) with a target cell discovery of 10,000 cells.  Single-cell RNA-Seq libraries were prepared for the 10x scRNA sequencing platform according to the manufacturer's instructions with the Chromium Next GEM Single Cell 3@ Reagent Kit v3.1 (Dual Index; 10x Genomics) with a target cell discovery of 10,000 cells.