GSM8285563: E_1_APLNrKO_ML233_3h_1; Danio rerio; RNA-Seq
Embryos were homogenized in Trizol solution (Invitrogen) and total RNA was purified using chloroform (Fischer) and then isopropanol (Fischer). RNA (1 μg) was reverse-transcribed using an oligo-dT primer and RevertAid kit (Thermo-Scientific). Total RNA samples for bulkRNAseq analysis were prepared with ~50 embryos/sample. Each experimental condition has been performed using biological quadruplicates to ensure reproducibility. Trizol (Invitrogen) was used to homogenize cellular extracts with the TissueLyzer (Qiagen). RNA purification was performed as described above. RNA was quantified and sent to Novogene  for Illumina library preparation and paired-end sequencing. Messenger RNA was purified from total RNA using poly-T oligo-attachedmagnetic beads. After fragmentation, the first strand cDNA was synthesizedusing random hexamer primers, followed by the second strand cDNAsynthesis using dUTP.