title: Illumina NovaSeq 6000 paired end sequencing; RNAseq (olfactory bulb) of temperature- and Franciscella challenged Atlantic cod (Gadhus morhua)
design: RNAseq (olfactory bulb) of temperature- and Franciscella challenged Atlantic cod (Gadhus morhua)
library_name: Ne23_p
library_construction_protocol: Samples were collected 1) the day before the temperature was increased, 2) when the water temperature reached 17oC and 3) 5 weeks post infection. Fish (n=6, length = 23±2cm; weight =114±33g) were dip netted from the tank and were euthanised with an overdose of Benzoak Vet. The olfactory organ was exposed by opening the olfactory cavity via the nostril. The right olfactory organ was immediately placed in BiopSafe containing 10% formalin (Mermaid Medical, Vedbæk, Denmark) for histological processing. On the other hand, the left olfactory organ was placed in RNAlater® (Ambion, Inc., Connecticut, USA), allowed tissue penetration at 4oC overnight and stored at -40oC prior to RNA isolation.  Total RNA was isolated from the olfactory organ using the Agencourt RNAdvance™ Tissue Total RNA Purification Kit (Beckman Coulter Inc., CA, USA). The purified RNA was quantified using NanoDrop 8000 Spectrophotometer (ThermoFisher Scientific, United States) and integrity was further assessed using the Agilent® 2100 Bioanalyzer™ RNA 6000 Nano Kit (Agilent Technology Inc., Santa Clara, CA, USA). All samples had an RNA Integrity Number (RIN) above 7.0. Total RNA samples were sent to the Norwegian Sequecing Center (OUS, Norway). The strand-specific TruSeq RNA Library Prep Kit (Illumina, CA, USA) was used for preparation of RNA-seq libraries, using the manufacturer's protocol.