The 4th Vienna Next Generation Sequencing Symposium & Workshop brought together over 150 current and future users of next generation sequencing technology. Six invited speakers described exciting applications of NGS technology in basic biology and in biomedical research. And the keynote speaker Bart Deplancke (École Polytechnique Fédérale de Lausanne, EPFL) presented a biologically interesting and methodologically advanced study on the massive-scale characterization of the genome regulatory code in Drosophila. The meeting was organized by Christoph Bock (Coordinator of the BSF Biomedical Sequencing Facility of CeMM and MedUni Vienna) and Andreas Sommer (Coordinator of the CSF NGS unit at the Vienna Biocenter), who also gave an overview of the technological support and opportunities for collaboration that are available to researchers in Vienna, Austria and beyond. The symposium was followed by an interactive workshop that allowed all participants to go into further detail and to discuss practical approaches and challenges of using NGS technology in their research. The success story of the Vienna NGS Symposium & Workshop will continue with 5th event in winter 2015/2016.