Fluidic larger-scale integration (LSI) resides at the heart of comprehensive sample-to-answer automation and parallelization of assay panels for frequent and ubiquitous bioanalytical testing in decentralized point-of-use/point-of-care settings. This paper develops a novel “digital twin” strategy with an emphasis on rotational, centrifugo-pneumatic flow control. The underlying model systematically connects retention rates of rotationally actuated valves as a key element of LSI to experimental input parameters; for the first time, the concept of band widths in frequency space as the decisive quantity characterizing operational robustness is introduced, a set of quantitative performance metrics guiding algorithmic optimization of disc layouts is defined, and the engineering principles of advanced, logical flow control and timing are elucidated. Overall, the digital twin enables efficient design for automating multiplexed bioassay protocols on such “Lab-on-a-Disc” (LoaD) systems featuring high packing density, reliability, configurability, modularity, and manufacturability to eventually minimize cost, time, and risk of development and production