A semi-automated approach using a liquid handling robot was developed to apply the luciferase-based assay in a highly accurate substrate screening assay for nucleoside kinases (Figure ). Scripts were written to pipette the kinase reaction by the robot into 96-well PCR plates. Afterwards, the plates were manually transferred to an incubator. After completion of the reaction, plates were manually transferred back to the deck of the robot and the luciferase reaction was pipetted. Subsequently, the assay plate was automatically transported to the plate reader to measure luminescence after 10 min incubation in the dark. Please see Table and the externally hosted Supplementary Information [34] for more detailed information on the approach.