GSM7067527: KO, replicate 1, scRNAseq; Danio rerio; RNA-Seq
The tissues of head regions from zebrafish embryos were washed in a phosphate-buffered solution before adding dispase I (Sigma, D4818) and incubated at 0.5U, 33 °C for 10 min, with 750 rpm shock (pipette every 30 s). After adding 100 μL of serum to stop digestion, tissues were filtered through a 70 μm mesh and a 20 μm mesh cell strainer, pelleted by centrifugation at 4 °C, and washed twice with ice-cold HBSS adding 0.5 % BSA to remove debris. Single cell suspensions were processed for library construction following the manufacturer's instructions (Chromium Next GEM Single Cell 30 Reagent Kits v3.1 User Guide). Libraries were then sequenced by Illumina Novaseq 6000 platform.