Noncontact Infrared-Mediated Thermocycling for Effective Polymerase Chain Reaction Amplification of DNA in Nanoliter Volumes
- 23 September 2000
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 72 (21) , 5507-5512
- https://doi.org/10.1021/ac000423j
Abstract
We demonstrate that accurate thermocycling of nanoliter volumes is possible using infrared-mediated temperature control. Thermocycling in the presence of Taq polymerase and the appropriate primers for amplification of λ-DNA in a total volume of 160 nL is shown to result in the successful amplification of a 500-base pair fragment of λ-DNA. The efficiency of the amplification is sufficiently high so that as few as 10 cycles were required to amplify an adequate mass of DNA for analysis by capillary electrophoresis. This indicates that, as expected, PCR amplification of DNA in nanoliter volumes should not only require less Taq polymerase but require less cycling time to produce a detectable amount of product. This sets the stage for microchip integration of the PCR process in the nanoliter volumes routinely manipulated in electrophoretic microchips.Keywords
This publication has 6 references indexed in Scilit:
- Degenerate Oligonucleotide Primed–Polymerase Chain Reaction and Capillary Electrophoretic Analysis of Human DNA on Microchip-Based DevicesAnalytical Biochemistry, 1998
- A Miniature Analytical Instrument for Nucleic Acids Based on Micromachined Silicon Reaction ChambersAnalytical Chemistry, 1998
- Fully Automated DNA Reaction and Analysis in a Fluidic Capillary InstrumentAnalytical Chemistry, 1997
- Micromachining a Miniaturized Capillary Electrophoresis-Based Chemical Analysis System on a ChipScience, 1993
- Minimizing the time required for DNA amplification by efficient heat transfer to small samplesAnalytical Biochemistry, 1990
- Automated polymerase chain reaction in capillary tubes with hot airNucleic Acids Research, 1989