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. 2014 Feb 18;9(2):e89331. doi: 10.1371/journal.pone.0089331

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© 2014 Nachar et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Pairwise variation (V_n/_n+1) calculated between the normalization factors NF_n and NF_n+1 to assess the optimal number of reference genes to use in normalisation. V<0.15 is considered sufficient to normalise. Here, the use of two to three genes have a V₂/₃ = 0.0575, which indicates that the use of two to three genes is sufficient to calculate the normalization factor.