title: GSM8033234: Ko-1; Danio rerio; RNA-Seq
library_construction_protocol: The 5 dpf larvae were anesthetized with 150 mg/L tricaine in groups of four at a time. The tip of the caudal fin of the anesthetized larvae was then removed using a hypodermic needle. The tail was transferred to a 96-well plate with 100% methanol-filled wells for genotyping. The larvae bodies were covered with 25 µL TRIzol (Invitrogen; #15596-018) and stored at -80°C until RNA extraction. According to the obtained genotyping results, bodies of hspd1+/+ and hspd1-/- larvae were pooled in groups of 5 larvae, and this process was repeated for 3 mating sets, resulting in 6 samples in total. Pooled samples were homogenized with a motorized pellet pestle (Kimble; #Z359971). RNA isolation was performed using TRIzol, including DNase treatment with DNA-free Kit (Ambion, AM1906) according to the manufacturer's instructions. Total RNA concentrations were determined with Synergy H1 plate reader equipped with Take3 Plate (BioTek, Agilent). Quality control analysis was performed with an automated 4200 TapeStation System (Agilent, G2991AA), and RNA integrity number (RIN) values were determined by RNA ScreenTape Assay for 4200 TapeStation System according to the manufacturer's instructions. RIN values were above 8.3 for all samples. RNA sequencing service was performed by BGI Copenhagen by a non-stranded and polyA-selected RNA library preparation and a consequent PE100 sequencing on DNBSEQ.immediately snap-frozen in an empty 96-well plate (Biorad; #12001925) on dry ice and stored at -80°C until sample preparation. Library preparation and RNA sequencing service was performed by BGI Copenhagen by a non-stranded and polyA-selected RNA library preparation and a consequent PE100 sequencing on DNBSEQ.