RqPCRAnalysis : analysis of quantitative Real-time PCR Data
Résumé
We propose the statistical RqPCRAnalysis tool for quantitative real-time PCR data analysis which includes the use of several normalization genes, biological as well as technical replicates and provides statistically validated results. This RqPCRAnalysis tool improved methods developed by Genorm and qBASE programs. The algorithm was developed in R language and is freely available. The main contributions of RqPCRAnalysis tool are: (1) determining the most stable reference genes (REF)-housekeeping genes-across biological replicates and technical replicates; (2) computing the normalization factor based on REF; (3) computing the normalized expression of the genes of interest (GOI), as well as rescaling the normalized expression across biological replicates; (4) comparing the level expression between samples across biological replicates via the test of statistical significance. In this paper we describe and demonstrate the available statistical functions for practical analysis of quantitative real-time PCR data. Our statistical RqPCRAnalysis tool is user-friendly and should help biologist with no prior formation in R programming to analyze their quantitative PCR data.