Research focuses on RNA-Seq analysis, to develop best practices and user-friendly workflows for processing and integrating gene expression data as well as to provide assistance for experiment design, analysis and interpretation. The variety of RNA-Seq study designs and characteristic properties of studied organisms greatly affect bioinformatics analysis. As part of the RNA Bioinformatics center (RBC), which supports all RNA-related research within de.NBI, de.STAIR summarizes the expertise of three participating laboratories to develop and maintain an flexible RNA-Seq analysis workbench for different RNA-Seq protocols, to be used locally or in a cloud environment as provided by the open source Galaxy platform for biomedical research. In addition, de.STAIR provides workshops, training programs and screencasts for experts and non-experts to lower the barriers to RNA-Seq data analysis.