Ferramenta (RGeasy®) de análise de genes de referência para estudos de expressão via RT-qPCR

Abstract

Gene expression through RT-qPCR can be performed by the relative quantification method, which requires the expression normalization through reference genes. Therefore, it is essential to validate, experimentally, the chosen reference genes. Thus, although there are several studies that are conducted to identify the most stable reference genes, most of these studies validate genes for very specific conditions, not exploring the whole potential of the research. It turns out that new experiments must be conducted for those that have interest in analyzing gene expression of treatments and/or conditions not explored in these studies. In this study, we present the software RGeasy, which aims to facilitate the selection of reference genes, allowing the user to choose genes for a higher number of combinations of treatments/conditions, compared to the ones present in the literature with just a few clicks. RGeasy was validated with data from gene expression studies performed in Coffea arabica and Coffea canephora, however it can provide information for all species from these three groups: animals, plants, and microorganisms. In addition to display a rank of the most stable reference genes for each condition or treatment, the user also has access to primer pairs for the selected reference genes.