RNA-seq of Atlantic salmon tongue and olfactory rosette -including infection trial
RNA-seq of Atlantic salmon tongue and olfactory rosette -including infection trial
Tongue samples for basal transcriptome profiling (RNA-seq) were collected from brackish water-adapted Atlantic salmon (~120 g, n=5) reared in a recirculating aquaculture system at 12 ppt salinity and 12°C. Fish were fasted for 24 hours before sample collection and euthanised using an overdose of Benzoak vet (ACD Pharmaceuticals AS, Norway). Tongue samples were preserved in RNAlater™ Stabilization Solution (Thermo Fisher Scientific, Lithuania), kept overnight at 4oC, and then transferred to -20oC until RNA isolation.    RNA of all samples was isolated using the Agencourt RNAdvance™ Tissue Total RNA Purification Kit (Beckman Coulter Inc., USA) using the Biomek 4000 automated workbench station (Beckman Coulter, Inc.). To provide an en masse representative of the organ, the entire tongue was homogenised first using ceramic beads in FastPrep-96™ (MP Biomedicals LLC, USA) for 2 mins before a small portion of the homogenate (ca 10 mg) was used for RNA isolation. RNA concentration and purity were assessed using a NanoDrop 8000 Spectrophotometer (Thermo Fisher Scientific). Samples designated for RNA sequencing underwent additional quality control using an Agilent 2100 Bioanalyzer (Agilent, California, USA). The RNA integrity number (RIN) values of all samples were above 8.0.   Total RNA samples were sent to the Norwegian Sequencing Centre (OUS, Norway). A strand-specific TruSeq RNA Library Prep Kit (Illumina, CA, USA) was used to prepare RNA-seq libraries using the manufacturer's protocol. 