GSM8704821: H9 DIV100 SpOrg - Sample A; Homo sapiens; RNA-Seq
Organoids were dissociated using papain (Worthington) on the gentleMACs octodissociator (Miltenyi; ABDK settings) Single cell libraries were prepared in accordance with the manufacturer using the Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1 (10X Genomics, 1000269) with a 10X Chromium Controller (10X Genomics, 1000204). In brief, the previously dissociated cell mixture and kit Mastermix was loaded onto a Chromium Single cell 3′ chip (estimated capture of 10,000 cells) to generate a gel beads in emulsion (GEM) mixture. A reverse transcriptase reaction was then carried out to generate a barcoded cDNA library using C1000 Touch Thermal Cycler (Biorad, 12 cycles of amplification). Quality control of samples was performed using a 2100 Bioanalyzer Instrument (Agilent) before further processing. For sequencing, samples were pooled and ran on an Illumina NovaSeq 6000.