RNA-Seq juvenile Scophthalmus maximus: head kidney
RNA was isolated from head kidney and intestine samples collected from fish exposed to OTC or vehicle alone and intraperitoneally injected with VHSV or cell culture at 24 hpt using the Maxwell RSC SimplyRNA Tissue Kit (Promega) on an automated Maxwell RSC 48 Instrument, following the manufacturers instructions. RNA concentration and purity were assessed using a Nanodrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA), and integrity was evaluated with an Agilent 2100 Bioanalyzer (Agilent Technologies, USA), according to the manufacturers guidelines. A total of 16 samples per tissue, four from each experimental group, were used for Illumina library preparation. Double-stranded cDNA libraries were constructed using the TruSeq Stranded mRNA LT Sample Kit (Illumina, San Diego, CA, USA). Paired-end 150 bp sequencing was performed on an Illumina NovaSeq X platform. Both library preparation and sequencing were carried out by Macrogen, Inc. (Seoul, Republic of Korea).