TY - GEN
T1 - A quantitative free convection DNA amplifier
AU - Daly, John
AU - Davies, Mark
PY - 2007
Y1 - 2007
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=43449110127&partnerID=8YFLogxK
U2 - 10.1115/HT2007-32381
DO - 10.1115/HT2007-32381
M3 - Conference contribution
AN - SCOPUS:43449110127
SN - 0791842746
SN - 9780791842744
T3 - 2007 Proceedings of the ASME/JSME Thermal Engineering Summer Heat Transfer Conference - HT 2007
SP - 41
EP - 50
BT - 2007 Proceedings of the ASME/JSME Thermal Engineering Summer Heat Transfer Conference - HT 2007
T2 - 2007 ASME/JSME Thermal Engineering Summer Heat Transfer Conference, HT 2007
Y2 - 8 July 2007 through 12 July 2007
ER -