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Determination of optimal number of reference genes based on pair-wise variation (V) analysis of normalisation factors of the candidate reference genes. A Vn/n+1 value is shown for every comparison between two consecutive candidate reference genes. As a general guideline it is stated that the benefit of using an extra (n+1)th reference gene is limited as soon as the Vn/n+1 value drops below the 0.15 threshold.
