title: GSM6792818: line 1, D115, organoid; Homo sapiens; RNA-Seq
library_construction_protocol: Approximately ten representative organoids were selected from culture. Organoids were allowed to settle by gravity in a 1.5 mL tube and medium was aspirated. Organoids were then dissociated with 20 units/mL of papain (Worthington Biochemical Corporation, Lakewood, NJ) with 60 units/mL DNase I (Worthington Biochemical Corporation) on a shaker at 37˚C for approximately 60 minutes. Following dissociation, cells were pelleted at 400 x g for 5 minutes, resuspended in dPBS -/- (Thermo Fisher Scientific) with 0.04% non-acetylated bovine serum albumin (New England Biolabs, Ipswich, MA), and processed immediately for encapsulation and barcoding using the Chromium Controller (10X Genomics, Pleasanton, CA). Cells were filtered through a 70 µm filter and further diluted to target 8,000 cells per run. Single cells were then partitioned and barcoded with the Chromium Controller instrument (10X Genomics) and Single Cell 3' Reagent (v3.1 chemistry) kit (10X Genomics) according to the manufacturer's specifications with no modification (Rev C). Final libraries were quantified using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific) and diluted to 3 ng/µL in buffer EB (Qiagen, Hilden, Germany). Library quality and concentration was confirmed using the Bioanalyzer High Sensitivity DNA Assay (Agilent Technologies, Santa Clara, CA) prior to sequencing. Libraries were prepared using the Single Cell 3' Reagent (v3.1 chemistry) kit (10X Genomics) according to the manufacturer's specifications with no modification (Rev C).