GSM5353448: runx1 homozygous [ko2]; Danio rerio; RNA-Seq
After euthanasia, the adult zebrafish were subjected to cardiac bleeding to remove most mature erythrocytes from the marrow. The zebrafish were then dissected at room temperature and the dissected marrow was placed into 500ul of 0.9x PBS with 2% FBS and 1 USP units/mL heparin, placed on ice. The marrow samples were then mechanically dissociated by repeated pipetting into single cell suspension and passed through a 40-μm nylon mesh. All 8 samples were washed by spinning down at 350 x g for 5 min at 4C. The cell pellets were collected in 300ul of 1x DPBS with 0.04% BSA (Gemini Bio-Products), then washed again after spinning down at 350 x g for 5 minutes at 4°C and counted after resuspension in DPBS/BSA to target 6,000 cells per reaction. We followed the 10x Genomics sample preparation protocol and the 10x Genomics single cell RNA sequencing v3.1 library preparation.