GSM5070576: npas4l_mut; Danio rerio; RNA-Seq
Tails from ~150 npas4l heterozygous (npas4lbns423/+) and trans-heterozygous (npas4lbns423/bns297) mutant embryos were cut at the anterior end of the yolk extension at 20 hpf in DMEM/F10+5%FBS+0.01% tricaine on agarose-coated plates. The cells were dissociated using the Pierce Cardiomyocyte dissociation kit according to manufacturer instructions and FACS sorted for GFP-fluorescence using an AriaIII-sorter and DAPI as indicator for dead cells. The cell suspensions were counted with Moxi cell counter and diluted according to manufacturer's protocol to obtain 5000 single cell data points per sample. Each sample was run separately on a lane in Chromium controller with Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (10xGenomics). Single cell RNAseq library preparation was done using standard protocol.