RNA-Seq of turbot intestine
RNA was isolated using the Maxwell RSC simplyRNA Tissue Kit (Promega) with an automated Maxwell RSC 48 Instrument, following the manufacturers instructions. RNA concentration was determined using a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA), and RNA integrity was assessed with an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturers protocol prior to library preparation for Illumina sequencing. The five RNA samples from each experimental group showing the highest quality were selected for transcriptome sequencing. TruSeq Stranded mRNA LT Sample Preparation Kit from Illumina was used for RNA sequencing, according to the manufacturer's instructions. A cDNA library was prepared from mRNA via reverse transcription. The amount of cDNA in each library was quantified through spectrofluorometric analysis using the Qubit system (Thermo Fisher Scientific, Massachusetts, United States). The paired-end 150 bp reads were obtained by Illumina NovaSeq X sequencer at Macrogen Inc. (Seoul, Republic of Korea).