Ultrafast, low-power, PCB manufacturable, continuous-flow microdevice for DNA amplification
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Checksum (MD5)
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Author(s)
Kaprou, Georgia D
Papadopoulos, Vasileios
Papageorgiou, Dimitris P
Kefala, Ioanna
Papadakis, George
Gizeli, Electra
Chatzandroulis, Stavros
Kokkoris, George
Tserepi, Angeliki
Date Issued
June 3, 2019
Journal
Analytical and Bioanalytical Chemistry
Publisher
Springer Berlin Heidelberg
Citation
Kaprou, Georgia D. et al. "Ultrafast, low-power, PCB manufacturable, continuous-flow microdevice for DNA amplification." Analytical and Bioanalytical Chemistry 411 (June 2019): 5297-307 ©2019 Author(s)
Version
Author's final manuscript
Abstract
The design and fabrication of a continuous-flow μPCR device with very short amplification time and low power consumption are presented. Commercially available, 4-layer printed circuit board (PCB) substrates are employed, with in-house designed yet industrially manufactured embedded Cu micro-resistive heaters lying at very close distance from the microfluidic network, where DNA amplification takes place. The 1.9-m-long microchannel in combination with desirably high flow velocities (for fast amplification) challenged the robustness of the sealing that was overcome with the development of a novel bonding method rendering the microdevice robust even at extreme pressure drops (12 bars). The proposed fabrication methods are PCB compatible, allowing for mass and reliable production of the μPCR device in the established PCB industry. The μPCR chip was successfully validated during the amplification of two different DNA fragments (and with different target DNA copies) corresponding to the exon 20 of the BRCA1 gene, and to the plasmid pBR322, a commonly used cloning vector in E. coli. Successful DNA amplification was demonstrated at total reaction times down to 2 min, with a power consumption of 2.7 W, rendering the presented μPCR one of the fastest and lowest power-consuming devices, suitable for implementation in low-resource settings. Detailed numerical calculations of the DNA residence time distributions, within an acceptable temperature range for denaturation, annealing, and extension, performed for the first time in the literature, provide useful information regarding the actual on-chip PCR protocol and justify the maximum volumetric flow rate for successful DNA amplification. The calculations indicate that the shortest amplification time is achieved when the device is operated at its enzyme kinetic limit (i.e., extension rate).
MIT Department
Massachusetts Institute of Technology. Department of Materials Science and Engineering
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1007/s00216-019-01911-1
