4.8.3. Cycling parameters

The default incubation times recommended for particular kits have been optimized for the reaction components and should be followed unless there are compelling reasons not to. Typical for PCR products <400 bp is 10 seconds denaturation at 95oC; 15 seconds annealing-extension at 58-60oC. Longer products require an additional incubation of 60 seconds per 1,000 bp at 72oC.