GSM8607835: Adult HdrR skin & scales, run2; Oryzias latipes; RNA-Seq
Fish were anesthetized with Tricaine and euthanized on ice. Skin and scales were removed using tweezers and immersed in TrypLE Express (Invitrogen, 12604039). The sample was incubated at 37°C and shaken at 300 rpm for 1 hr, followed by mechanical disruption with a the isolated cells were incubated with Draq5 (Thermo Scientific™, Cat. 62251) to mark the nuclei for 10 min prior to FACS. Cells were analyzed and sorted using a BD FACSAria II Special Order Research Product (SORP) Cell Sorter with a 100 µM nozzle. Cells were counted and diluted to a final concentration in PBS supplemented with 0.04% BSA. Cellular suspension was loaded onto Next GEM Chip G targeting first experiment 10,000 cells and second experiment 1,000 cells, then ran on a Chromium X instrument to generate gel beads in emulsion (GEMs, 10x Genomics). Single-cell 3' RNA-seq libraries were generated according to the manufacturer's protocol (10x Genomics Chromium Single Cell 3' Reagent Kit User Guide v3.1 Chemistry). : Single-cell 3' RNA-seq libraries were quantified using NEBNext Library Quant Kit for Illumina (NEB) and high sensitivity D1000 TapeStation (Agilent). Libraries were pooled according to targeted cell number, aiming for ~25,000 reads per cell. Pooled libraries were sequenced on a NovaSeq 6000 instrument using an SP 100 cycles reagent kit (Illumina) with the following read length: 28 bp Read1, 10 bp I7 Index, 10 bp I5 Index and 90 bp Read2. Single-cell 3' RNA-seq
GEO
GSM8607835_r1