GSM8146664: B.gargarizans_PG, ScRNAseq; Bufo gargarizans; RNA-Seq
Adult toad postauricular glandular tissue was used, cut into approximately 1 mm pieces with sterilized scissors at room temperature, transferred into 15 ml centrifuge tubes, washed with PBS, and suspended in 3-4 ml of solution (TrypLE (1 mg /ml) containing collagenase I) and digested for 60 min until no tissue fragments were visible. To collect free cells, the tubes were centrifuged at 600 × g for 5 min at 4°C, the supernatant was removed and resuspended in 2 ml PBS. The dissociated cells were then passed through a 70 µm filter (Cat. No. 10) and then through a 40 µm filter (Cat. No. 10). After washing 2 times (centrifugation at 600 × g for 5 min at 4°C), they were resuspended in DPBS at a density of 1 × 105 cells/ml. Generation of single-cell RNA sequencing libraries and signature barcode matrices. library was performed according to the manufacter's instructing (Single Cell 3' V3.1, 10x genomices)