RNA-seq salmo salar head-kidney
Libraries for the fish from each tank were prepared using the TruSeq RNA Library Preparation Kit v2 Set A (Cat N  RS1222001, Illumina Inc., San Diego, CA, USA) according to the manufacturer s instructions (Cat. N  RS1229004DOC, Part N  15031047 Rev. E). The libraries were analysed in a High Sensitivity Bioanalyzer 2100 with chips (Agilent Technologies, Santa Clara, CA, USA) using the KAPA Library Quantification kit (Cat. N  KK4824 Version 4.11, Kapa Biosystems, Wilmington, MA, USA) according to the manufacturer s instructions. The libraries were diluted to 10 nM in 10 mM Tris-HCl buffer (pH 8.5) containing 0.1% Tween 20. Then, the libraries were diluted to 4 nM according to the manufacturer s instructions (Cat N  RS1222001, Illumina Inc., San Diego, CA, USA). For sequencing, 12.5 pM RNA from the pool and 1% PhiX were loaded into the Illumina MiSeq sequencer (Illumina Inc., San Diego, CA, USA) and sequenced using the MiSeq Reagents v3 600 cycles sequencing kit (Cat. N  MS1023003, Illumina Inc., San Diego, CA, USA). A quality check of the libraries was performed by classification and quantification of the PCR products using the 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) and the Illumina/Universal library quantification kit (Kapa Biosystems, Wilmington, MA, USA) according to the manufacturer s instructions.