Compatibility of Segmenting Fluids in Continuous-Flow Microfluidic PCR

[+] Author and Article Information
E. J. Walsh

Stokes Research Institute,  University of Limerick, Castletroy, Limerick, Irelandedmond.walsh@ul.ie

C. King, R. Grimes, A. Gonzalez, D. Ciobanu

Stokes Research Institute,  University of Limerick, Castletroy, Limerick, Ireland

J. Med. Devices 1(4), 241-245 (Sep 12, 2007) (5 pages) doi:10.1115/1.2812426 History: Received February 20, 2007; Revised September 12, 2007

Continuous flow offers notable advantages over batch processing for analytical applications like gene expression profiling of biological material, which demands very high processing. The technology of choice for future genetic analyzers will most likely use the polymerase chain reaction (PCR); therefore, high-throughput, high-speed PCR devices have raised enormous interest. Continuous-flow, biphasic PCR can meet these requirements but segmenting∕carrier fluids chemically compatible with the PCR are needed. The present paper compares several fluids in terms of compatibility with PCR and fluidic dynamics in a continuous, two-phase flow microfluidic device, and PCR efficiency was assessed quantitatively. The results represent the first step toward rational fluid design for biphasic continuous PCR.

Copyright © 2007 by American Society of Mechanical Engineers
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Figure 1

Isometric (top) and end view (bottom) of PCR device utilized to both generate segmented flows and perform the PCR during the course of the present experimentation. Dimensions are in millimeters.

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Figure 2

PCR performance in samples amplified in AB7900HT in the presence of 4μl of overlaid oil

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Figure 3

Triplicate dissociation curves for PCR amplified in the presence of (a) FC40 plus surfactant or (b) AR20 silicone oil as segmenting fluid

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Figure 4

Mean F and dF∕dT data as a percentage of the mean of the positive controls

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Figure 5

Flow visualization showing the film thickness between an aqueous plug and the tubing wall for the potential segmenting fluids under investigation. Average flow velocity is 100mm∕s.




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