GSM8521356: Inner ear organoid d40 SOX2-ntdT+ Matrigel; Homo sapiens; RNA-Seq
Aggregates were collected in a 2 mL tube, centrifuged (200 x g for 3 min), and treated with Accutase at 37 C for 90 min. Aggregates were periodically, gently pipetted with p1000 tips to mechanically dissociate. After confirming dissociation, aggregates were sequentially filtered through a 100 µm and 40 µm cell strainer. Aggregates were then transferred into a 2 mL tube, centrifuged (100 x g for 5 min), and resuspended with fluorescence-activated cell sorting (FACS) buffer containing 2% fetal bovine serum in DPBS. SOX2ntdTomato-positive cells were collected via FACS. Library preparation was conducted using the 10X Genomics Chromium X platform with the 10X 3' scRNA-seq standard assay v3.1. The resultant cDNA library was sequenced with an Illumina NovaSeq X Plus system. Illumina's CellRanger v4.0.0 program was used to generate mapped, filtered gene-barcode matrices.