Recently, digital microfluidic biochips (DMFBs) have revolutionized many biochemical laboratory procedures and received much attention due to many advantages such as high throughput, automatic control, and low cost. To meet the challenges of increasing design complexity, computer-aideddesign (CAD) tools have been involved to build DMFBs efficiently. Current CAD tools generally conduct a twostage based design flow of fluidic-level synthesis followed by chip-level design to optimize fluidic behaviors and chip architecture separately. Nevertheless, existing fluidic-chip design gap will become even wider with a rapid escalation in the number of assay operations incorporated into a single DMFB. As more and more large-scale assay protocols are delivered in current emerging marketplace, this problem may potentially restrict the effectiveness and feasibility of the entire DMFB realization and thus needs to be solved quickly. In this paper, we propose the first fluidicchip co-design methodology for DMFBs to effectively bridge the fluidic-chip design gap. Our work provides a comprehensive integration throughout fluidic-operation scheduling, chip layout generation, control pin assignment, and wiring solution to achieve higher design performance and feasibility. Experimental results show the effectiveness, robustness, and scalability of our co-design methodology on a set of real-life assay applications.