title: RNA-Seq of Zebrafish larvae
design: Total RNA was isolated and purified from zebrafish roe tissues using TRIzol reagent.RNA quantity and purity were quantified using an ultra-micro spectrophotometer, and agarose gel electrophoresis was used to avoid alteration of RNA integrity and purity. RNA and DNA were quantified using the Qubit 2.0 RNA and Qubit 2.0 DNA kits. Subsequently, the extracted RNA was transformed to construct cDNA sequencing libraries using the TrueLib mRNA Library Prep Kit for Illumina. After quantification and pooling of cDNA using AgilentBioAnalyzer 2100, sequencing was performed on NovaSeq 6000. The quality of sequencing data was assessed and low quality sequences were filtered by FastQC (0.11.9) and Trimmomatic (0.32). Filtered sequenced sequences were aligned with the zebrafish reference genome using HiSAT2 (2.2.1), and finally transcripts were analyzed for gene expression levels by StringTie software