Gene Annealing Temperature (°C) Standard Curve Efficiencies Protocol Interleukin 1β 55 0.91 3 Step Interleukin 12 55 0.93-1.04 3 Step Immunoglobulin T 55 0.93-0.95 3 Step Matrix Metalloproteinase 9 55 0.90-0.91 3 Step Interleukin 8 55 0.91-0.93 3 Step Elongation Factor 1 55 0.91-0.93 3 Step Elongation Initiation Factor 55 1-1.06 3 Step Ribosomal Protein S20 55 0.93-0.95 3 Step

Table 3: Quantitative PCR protocols used to amplify reference genes and genes of interest in Atlantic salmon. Three step protocols consist of: One cycle of 95°C for 10 minutes, forty cycles of 95°C for 15 seconds, TAnnealing°C for 15 seconds and 72°C for 15 seconds, and one melt curve.

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