Abstract

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 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.

Highlights

  • Microfluidic systems have attained enormous recognition in the last two decades due to their characteristics of shortening the reaction time and fast transition within the process

  • Substrate-Cover Material In CF-polymerase chain reaction (PCR) system, silicon, glass, PDMS or hybrid stack are commonly utilized as substrate In continuous-flow PCR (CF-PCR)

  • The formation patterns of the temperature gradients are numerically validated for the various protrusion lengths of the invar sheet from the heater, which showed a very good agreement with experimental data, to enable a successful temperature target zone using a single heater with reliable target specificity

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Summary

Introduction

Microfluidic systems have attained enormous recognition in the last two decades due to their characteristics of shortening the reaction time and fast transition within the process. Microfluidic chips have made significant contributions to many biomedical and genetic related research fields. Polymerase chain reaction (PCR) is a biomedical technique with great potential for on-site evidence collection system of various pathology and food samples. With the invention of polymerase chain reaction (PCR) in 1986 [1], an in-vitro enzymatic amplification of nucleic acid, it has become one of the most appropriate regimes applicable on the microscale biochemical reactions. PCR is one of the most commonly used enzymatic processes that involve discrete thermal cycling, making it a good option to study heat transport characteristics in microfluidic chips. Thermal modeling is a key Processes 2019, 7, 919; doi:10.3390/pr7120919 www.mdpi.com/journal/processes

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