GSM6482080: wild-type  zebrafish embryos RNA-seq 72 hpf; Danio rerio; RNA-Seq
For bulk RNA-seq, RNA libraries were prepared for sequencing using standard Illumina protocols. For Ribo-seq, WT and mycn mutant embryos were collected at 3 dpf, digested with cold 0.5% trypsin to a single-cell suspension, and homogenized in lysis buffer for 20–50 times. The homogenate centrifuged at 12,000 g for 15 min at 4°C, and the supernatants were digested with RNase for 30 mins and aborted by 1M EGTA followed by layering on top of a 10%–50% sucrose-gradient solution. Ultracentrifugation was performed at 36,000 rpm for 2 hrs (hours) at 4°C. After centrifugation, the fractions were collected according to the absorbance at optical density OD260 by a TRAIX detector. Then the RNA was purified and used to construct the library for sequencing.