GSM5655700: Uninjected; Danio rerio; RNA-Seq
Embryos were treated with a dissociation mix consisting of 480 ml 0.25% trypsin-EDTA + 20 ml Collagenase 100 mg/ml for 15 mins at 30C. Once embryos were fully homogenized, dissociation was stopped with addition of 800 ml of DMEM-10%FBS. The homogenate was run though a 70 uM strainer before centrifugation for 5 min at 700 g at room temperature. The pellet was resuspended in 1mL PBS and centrifuged again for 5 min at 700 g at room temperature. The pellet was resuspended in 500uL PBS + 2% FBS and run through a 70uM cell strainer the strainer was then washed with 500uL of PBS + 2% FBS giving a total volume of 1mL. Libraries were prepared for sequencing using Illumina Single Cell 3' v3.1 Reagents under standard conditions scRNA-seq