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. 2011 Feb 9;4:36. doi: 10.1186/1756-0500-4-36

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Copyright ©2011 Piek et al; licensee BioMed Central Ltd.

This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Pairwise variations between 2 sequential normalization factors including an increasing number of potential reference genes . To determine the optimal number of reference genes, first the geometric mean of the expression of the previously ranked genes was calculated and then pair wise variations between sequential normalisation factors were calculated. Using the cut-off recommended by GeNorm of 0.15 the optimal number of reference genes for the data set in this study would be at least 5.