GSM8834936: gpr85-expressing cells from adult zebrafish Gpr85-Ctl brain; Danio rerio; RNA-Seq
Adult zebrafish brains were dissected and digested with 14 U papain (Sigma-Aldrich, p4762) at 33°C for 10 to 15 minutes in 0.9X Dulbeccoʹs Phosphate Buffered Saline (DPBS). Cell suspensions were washed and centrifuged (300 g) in 2% fetal bovine serum diluted in 0.9X DPBS (FACS buffer). Then, cells were resuspended in FACS buffer and filtered using a sterile 40 µm nylon mesh (VWR). Cell sorting of eGFP+ cells was performed on a FACS ARIA (Becton Dickinson, San Jose, CA) using Sytox Red (5 nM, Thermofisher scientific S34859) in order to remove dead cells. 50,000 cells were sorted per condition with an expected final density of 294 cells/µL. The library was prepared following 10x Genomics Chromium Single cell 3' kit (v3) guidelines.
GEO
GSM8834936_r1