GSM8553897: GFP+ TKO 4 dpf; Danio rerio; RNA-Seq
WT and TKO embryos were collected at 4 dpf, and GFP+ or dsRED+ cells from dissociated embryos were were isolated by FACS. For GFP+ cells, a manual resection of the tail and the head of each embryo was performed to enrich the pancreatic population of GFP+ cells. Single cell suspensions were generated using a previously published protocol (DOI: 10.1016/j.mex.2018.10.009). 10,000 cells were targeted from each sample cell suspension. The cells were washed twice with PBS+0.04% BSA and resuspended in about 500 cells per microliter. Single cells were captured using 10x Genomics Chromium. 10X Genomics's Chromium instrument and Dual index Single-Cell 3′ Reagent kit (V3.1) were used to prepare the individually barcoded single-cell RNA-Seq libraries following the manufacturer's protocol. Libraries quality was assessed by the TapeStation-4200 traces (Agilent BioAnalyzer High Sensitivity Kit) and quantitated by the Qubit system.