RNA-seq of Cyprinus carpio var. Koi: the whole fish  in 44 dph
TRIzol regent (Invitrogen, Carlsbad, USA) was used to extract total RNA. Then, the quality and integrity of RNA were verified by NanoDrop (Thermo Scientific, USA) and Agilent 2100 Bioanalyzer (Agilent Technologies , USA), respectively. Equal amounts of high quality RNA from 10-15 individuals in the same developmental stages were pooled and purified to construct gene expression profiling. Reverse transcription followed by PCR was applied to synthesize cDNA. Subsequently, the amplified cDNA fragments were purified and prepared for sequencing analysis using Illumina HiSeq 2500