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Figure 2 | BMC Plant Biology

Figure 2

From: Validation of reference genes for quantitative real-time PCR during leaf and flower development in Petunia hybrida

Figure 2

Data analysis flow chart. CT (cycle threshold) values were calculated using different thresholds depending on the variety. Efficiency value taken for line Mitchell was 2; for line V30, there was one value for each tube. Circles indicate statistical results to be merged with RankAggreg (Pihur 2009). Relative quantities (RQ) were scaled to the sample with lowest CT value (flower stage C). CT data were checked for normality (Shapiro-Wilk test) and, due to non-normality, they were analysed by non-parametrical tests (Kruskal and Wallis). Since CT values showed non-equal distributions according to the organ from which RNA was extracted, they were further tested using pairwise Wilcoxon tests with Bonferroni's correction with the aim of solving pairwise significant variations. A significance threshold of 0.05 was used. Abbreviations: PV, pairwise variation; M, classical stability value; stab, NormFinder stability value; CV, variation coefficient; r2, determination coefficient - regression to BestKeeper; RQ, relative quantities.

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