To respond to the dire need for miniaturization and process simplification of continuous-flow PCR (CF-PCR) device, this paper represents design and operation guide of a novel metal alloy assisted hybrid microdevice (polydimethylsiloxane (PDMS) and glass) for CF-PCR employing one heater. In this research, the specific objectives are to determine whether one heater chip design will be flexible enough when the size of DNA base pair is varied and to investigate whether one heater CF-PCR device will

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... be able to resolve the longstanding problem of thermal crosstalk. Furthermore, the parametric study is performed to determine which of the fourteen parameters have the greatest impact on the performance of one heater CF-PCR device. The main objective of this parametric study is to distinguish between the parameters that are either critical to the chip performance or can be freely specified. It is found that substrate thickness, flow rate, channel spacing, aspect ratio, channel pass length and external heat transfer coefficient are the most limiting parameters that can either improve or deteriorate the chip's thermal performance. Overall, the impact of design and operating parameters are observed to be least on thermocycling profile at low Reynolds number (≤0.37 Re). However, in addition to the primary metric advantages of CF-PCR, one heater chip design helps in minimizing the thermal crosstalk effects by a factor of 4 in comparison to dual heater PCR while still maintaining a critical criteria of chip flexibility in terms of handling various sizes of DNA fragments. Hence, the proposed scheme paves the way for low-cost point-of-care diagnostics, system integration, and device miniaturization, realizing a portable microfluidic device applicable for on-site and direct field uses.