Noncontact Infrared-Mediated Thermocycling for Effective Polymerase Chain Reaction Amplification of DNA in Nanoliter Volumes

摘要

We demonstrate that accurate thermocycling of nanoliter volumes is possible using infrared-mediated temperature control. Thermocycling in the presence of Taq polymerase and the appropriat primers for amplification of #gamma#-DNA in a total volume of 160nL is shown to result in the successful amplification of a 500-base pair fragment of #gamma#-DNA. The efficiency of the amplification is sufficiently high so that as few as 10 cycles wer required to amplify an adequate mass of DNA for analysis by capillary electrophoresis. This indicates that, as expected, PCR amplification of DNA din naoiliter volumes should not only require less Taq polymerase but require less cucling 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.