GSM6214560: Ver2; Homo sapiens; RNA-Seq
Organoids were cultured to 120 days, then washed in PBS and dissociated using the gentleMACS Dissociator (Miltenyi Biotec) in program NTDK1 using the enzyme mix: Trypsin (Sigma Aldrich)/Accutase (Sigma Aldrich) (1:1) containing 10 U/ml DNaseI (Thermo Fisher Scientific). The washed cell suspension was passed through a 70μm cell strainer. Cells were counted and the suspension was loaded onto a Chromium Single Cell 3′ B Chip (10x Genomics, PN-1000073) and processed through the Chromium controller to generate single-cell GEMs (Gel Beads in Emulsion). scRNA-seq libraries were prepared with the Chromium Single Cell 3′ Library & Gel Bead Kit v.3 (10x Genomics, PN-1000075). Ready 10X libraries were sequenced paired end (R1:28, R2: 89 cycles) on NovaSeq (Illumina).