GSM6523916: Adult_adb1-2; Danio rerio; RNA-Seq
Forebrains were dissected from wild-type fish in Neurobasal supplemented with 1X B27 and promptly dissociated with the Papain Dissociation Kit. The cells were dissociated by gentle trituration 20 times and spun at 300xg for 5 minutes, resuspended in papain inhibitor in EBSS, spun at 300xg for 5 minutes. The resulting pellet was then washed in Neurobasal supplemented with B27 before final resuspension in 50 µL PBS + 200 µg/mL BSA. To avoid cell loss during each wash, the supernatant was saved and spun a second time to recover the remaining cells. The cells were then mixed with the final suspension to increase cell count. Viability and cell number were assessed by trypan blue staining on 10ul of the sample. Library was performed according to the manufacter's instructions (single cell 3' v2 protocol, 10x Genomics).