Abstract Summary: KNIME (Konstanz Information Miner) is a user-friendly and comprehensive open-source data integration, processing, analysis and exploration platform. We present here new functionality and workflows that open the door to performing next-generation sequencing analysis using the KNIME framework. Availability: All sources and compiled code are available via the KNIME update mechanism. Example workflows and descriptions are available through http://tech.knime.org/community/next-generation-sequencing. Contact: bernd.jagla@pasteur.fr Supplementary Information: Supplementary data are available at Bioinformatics online.