Total RNA was was extracted from 5 dpf larvae using TRI Reagent (Invitrogen, Cat. No. AM9738). Thirty larvae were pooled together to obtain one RNA sample. Total RNA was digested by DNase I and purified using the RNA-Clean and Concentrator Kit (ZYMO Research, Irvine, CA, USA) as the manufacturer described. Quality of RNA was checked by measuring absorbance at 260, 280, and 230 nm and Bioanalyzer Nano RNA chip (Agilent, USA) analysis. Samples with 260/280 nm as well as 230/280 nm absorbance ratio>1.8 and RNA integrity number (RIN) values above eight were used for further processing. RNA libraries were prepared from 1 ug of total RNA, for each sample, using KAPA Stranded RNA-seq kit (KK8401 from Roche). Breafly, mRNA was isolated with poliA beads from Illumina. The 1st and 2nd strands of DNA were synthetise and A tailing and ligation reactions were run. In ligation step the TruseqLT RNA adapter (Illumina) were used. The libraies were amplified runing 8 cycle of PCR.Quantity of each library were measure with Quantus device, then libraies were pooled in ratio 1:1 and concentration of the pool was confirmed with real-time PCR using KAPA Library Quantification kit (Roche). Libraries were sequence as pair-end (2 x 75 bp) on NextSeq 500 Illumina platform.