Computational protocol: Normalizing to GADPH jeopardises correct quantification of gene expression in ovarian tumours – IPO8 and RPL4 are reliable reference genes

Similar protocols

Protocol publication

[…] Descriptive statistics, F-test for Ct variance equality and Kolmogorov-Smirnov test for normality of log-transformed relative expression values were calculated by software SPSS 19.0 (SPSS Inc, Chicago, IL). The Equivalence test [-] and statistical applets BestKeeper [], geNorm [], and NormFinder [] were used for analysis of genes expression stability. GeNorm calculates a gene-stability measure, M-value, as the average pair-wise variation of a particular gene to all other candidate reference genes []. On the other hand, the stability value calculated with NormFinder combines estimated both intra-group and inter-group variations []. Genes with the lowest M-values have the most stable expression (least variability). Relative expression values for target genes were analysed by Kruskal-Wallis and Mann–Whitney tests, and the log-transformed values by one-way ANOVA. P < 0.05 was considered significant. […]

Pipeline specifications

Software tools BestKeeper, NormFinder
Application qPCR
Diseases Neoplasms, Ovarian Neoplasms