Recent advances in flow-based microfluidic biochips have enabled the emergence of lab-on-a-chip devices for bimolecular recognition and point-of-care disease diagnostics. However, the adoption of flow-based biochips is hampered today by the lack of computer-aided design tools. Manual design procedures not only delay product development but they also inhibit the exploitation of the design complexity that is possible with current fabrication techniques. In this paper, we present the first practical problem formulation for automated control-layer design in flow-based microfluidic very large-scale integration (mVLSI) biochips and propose a systematic approach for solving this problem. Our goal is to find an efficient routing solution for control-layer design with a minimum number of control pins. The pressure-propagation delay, an intrinsic physical phenomenon in mVLSI biochips, is minimized in order to reduce the response time for valves, decrease the pattern set-up time, and synchronize valve actuation. Two fabricated flow-based devices and six synthetic benchmarks are used to evaluate the proposed optimization method. Compared with manual control-layer design and a baseline approach, the proposed approach leads to fewer control pins, better timing behavior, and shorter channel length in the control layer.