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

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Protocol publication

[…] Data analysis strategy is described in detail in results. Reaction efficiency calculus was done using the amplification curve fluorescence, analyzing each tube separately as described by Liu and Saint (2002) []. It was calculated as follows: Efficiency = F(n)/F(n-1), in which n is defined as the 20% value of the fluorescence at the maximum of the second derivative curve. Curve was defined by one measure in each amplification cycle. We used only the exponential phase of the amplification reaction. Software packages included geNorm v3.4, the excel add-in of NormFinder v0.953, BestKeeper v1 and qBasePlus v1.2. Other statistical procedures were performed with the R program http://www.R-project.org, v2.7.1 with the packages stats v2.7.1, multcompView v0.1-0 and RankAggreg v0.3-1[]. […]

Pipeline specifications

Software tools geNorm, NormFinder, BestKeeper, RankAggreg
Applications Gene expression microarray analysis, qPCR
Diseases Blood Platelet Disorders