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Fig. 3 | Plant Methods

Fig. 3

From: Reference gene identification for reliable normalisation of quantitative RT-PCR data in Setaria viridis

Fig. 3

Average expression stability (M), ranking and pairwise variation (V n /V(n+1)) of the eleven reference gene candidates as calculated by NormFinder (a) and geNorm (b, c). A reduced value for ‘average expression stability’ indicates that the candidate reference gene is more stably expressed across the assessed tissues. Both the Normfinder (a) and the geNorm (b) approach ranked PP2A, ASPR6 and DUSP as the most stably expressed of the eleven candidates analysed. c Pairwise variation (V n /V(n+1)) analysis by geNorm determines the optimal number of reference gene candidates required for the accurate and reliable normalisation of RT-qPCR generated data. The use of three reference genes to accurately and reliably normalise RT-qPCR generated data across S. viridis tissues was decided upon due to the V3/4 value being lower than the proposed threshold value of 0.15 [21]

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