GSM9694967: 3 dpf wild-type tail; Danio rerio; RNA-Seq
Tails were pooled for the pax9 mutant and wild-type samples, then dissociated into a single-cell suspension largely as described (Bresciani et al., 2018), with Liberase substituting for collagenase. Tail biopsies were washed in PBS, then incubated in a sterile dish with a pre-warmed dissociation mixture of Liberase and Trypsin-EDTA (40 μl/8% 25x Liberase: 460 μl Trypsin-EDTA) in a standard incubator set at 30°C with the plate placed directly on a heat block. Dissociation progress was monitored over 15-20 minutes, and gentle trituration was performed every 3 minutes through a P200 pipette tip against the bottom of the dish, being careful to avoid bubbles and aggressive pipetting. Once a single-cell suspension was achieved, a stop solution of Leibovitz's media containing 10% FBS was added to halt dissociation. Pellets were washed in PBS containing 1% fetal bovine serum, filtered, and resuspended in 50 µl of 1% FBS/PBS. Samples were kept on ice for transfer to the core facility, where cell viability was confirmed at over 79%. Libraries were prepared using the Chromium GEM-X 3' Kit v4 according to the manufacturer's instructions
GEO
GSM9694967_r1