GSM4101381: 587680_46; Danio rerio; RNA-Seq
Hearts were extracted and transferred to Ringer solution with heparin sodium salt (50 U/ml) (Ringer composition: 7.2 g NaCl, 0.225g CaCl2.H2O, 0.37 g KCl, 0.2175g Na2HPO4.7H2O, 0.02g KH2PO4 at pH 7.4 sterilized by using a 0.22um bottle top filter unit) and rinsed once with Phosphate buffered saline (PBS) or directly isolated in PBS. Hearts were inspected, cleaned and then fixed with 4% Paraformaldehyde (PFA) overnight (O/N) on nutator at room temperature (RT). Samples were rinsed once in PBS, dehydrated into Ethanol (EtOH) at 70%, 80%, 90%, 96%  for 2 hours each step and 2x100% for 1 hour each step, followed by a 100% 1-butanol step overnight. The samples were then transferred to paraffin (Paraplast Plus, Sigma-Aldrich P3683) wax at 65°C. Cells were washed in PBS with 0.04% BSA and re-suspended before loading 12,000-12,500 cells onto each channel of the Chromium 10x Genomics platform to capture single cells in droplets. Library generation for 10x Genomics v2 chemistry was performed following the Chromium Single Cell 3ʹ Reagents Kits User Guide: CG00052. Quantification of cDNA was performed using Qubit dsDNA HS Assay Kit (Life Technologies Q32851) and high-sensitivity DNA tape-station (Agilent. 5067-5584). Quantification of library construction was performed using Qubit dsDNA HS Assay Kit (Life Technologies Q32851) and high-sensitivity DNA tape-station (Agilent. 5067-5584).
GEO Accession
GSM4101381