GSM3938460: Knockout Embryonic Day 18.5; Mus musculus; RNA-Seq
At noon, embryos were harvested by hysterectomy and the pancreas was dissected under a microscope in ice-cold phosphate buffered saline (PBS) and transferred into 2 mL of prewarmed 0.25% Trypsin-EDTA (TE) in a 15 mL Falcon tube. Falcon tubes were placed in a 37°C bead bath for 20 minutes of digestion, followed by addition of 1 mL of FBS and 2 mL of PBS to stop the digestion. The cell solution was passed through a 40 µM cell strainer into a new 15 mL Falcon tube. Single cells were spun at 300xg for 5 minutes and cell pellets were resuspended in 500 µL (control) or 250 µL (Dpy30∆P) PBS and cell concentration was adjusted to 1,000 cells/µL. Using a cell concentration of 1,000 cells/µL, 17.4 µL of cells were loaded for a targeted recovery of 10,000 cells according to the Chromium Single Cell 3' Reagent Kits v2 User Guide (10X Genomics, Inc.), which was followed as described. Single Cell RNA-seq