GSM8826834: CTRL, rep3; Homo sapiens; RNA-Seq
Organoids were dissociated using an optimized protocol based on the Worthington Papain Dissociation System Kit (Worthington Biochemical). Single cell suspensions were collected on ice-cold PBS (without calcium and magnesium) with 0.04% BSA at 1×10^6 /ml as counted by an automatic cell counter. Library was performend accordingly to manufacter's instructions (10X Genomics). Briefly, approximately 8000 cells from each sample prepared as described above were loaded into one channel of the Single Cell Chip B using the Single Cell 3' v3 reagent kit (10X Genomics) for gel bead emulsion generation in the Chromium system. Following capture and lysis, cDNA was synthesized and amplified for 14 cycles following the manufacturer's protocol (10X Genomics). The amplified cDNA (50 ng) was then used for each sample to construct Illumina sequencing libraries. Sequencing was performed on the NextSeq550 Illumina sequencing platform following the 10x Genomics instructions for read generation. A sequencing depth of ~45,000 reads/cell was obtained for each sample.