The Polymerase Chain Reaction (PCR) has been used extensively to amplify targeted nucleic acids for many applications in molecular biology and, increasingly, in medical diagnostics. Outlined in this paper is a PCR device which takes account of the advantages offered by free convection. The design is, in it fundamental format a time-wise isothermal well-based thermocycler. A temperature gradient induced across the well causes convection forces to circulate the sample through the required temperatures necessary for amplification. Quantitative amplification is demonstrated with real time measurements of SYBR Green I fluorescence within the free convective DNA amplifier. Amplification of an 86-bp fragment of the pGEM®-T vector (Promega) is performed in a 25μl volume in eight minutes. A 10-fold dilution series and methods for calculating effective cycle times are presented. Also detailed within this paper are PIV and thermal imaging results of the free convection cavity. This device presents an opportunity for the development of a practical and inexpensive gene-expression measurement system.
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ASME/JSME 2007 Thermal Engineering Heat Transfer Summer Conference collocated with the ASME 2007 InterPACK Conference
July 8–12, 2007
Vancouver, British Columbia, Canada
Conference Sponsors:
- Heat Transfer Division
ISBN:
0-7918-4276-2
PROCEEDINGS PAPER
A Quantitative Free Convection DNA Amplifier Available to Purchase
John Daly,
John Daly
University of Limerick, Limerick, Ireland
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Mark Davies
Mark Davies
University of Limerick, Limerick, Ireland
Search for other works by this author on:
John Daly
University of Limerick, Limerick, Ireland
Mark Davies
University of Limerick, Limerick, Ireland
Paper No:
HT2007-32381, pp. 41-50; 10 pages
Published Online:
August 24, 2009
Citation
Daly, J, & Davies, M. "A Quantitative Free Convection DNA Amplifier." Proceedings of the ASME/JSME 2007 Thermal Engineering Heat Transfer Summer Conference collocated with the ASME 2007 InterPACK Conference. ASME/JSME 2007 Thermal Engineering Heat Transfer Summer Conference, Volume 3. Vancouver, British Columbia, Canada. July 8–12, 2007. pp. 41-50. ASME. https://doi.org/10.1115/HT2007-32381
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