Development of a multireactor microfluidic system for the determination of DNA using real-time polymerase chain reaction

A microfluidic system that allowed us to perform the real-time polymerase chain reaction (PCR) in a glass-silicon microchip containing nine 250-nL microreactors was developed and studied. The resulting high heating/cooling rates of a PCR mixture in a microreactor allowed us to optimize the amplification mode (1 min/cycle). The silicon surface of microreactors was successfully passivated. The resulting analytical system allowed us to measure the PCR kinetic curves in chip microreactors at a DNA concentration of ～5 × 10⁴ copies per microreactor. It was found that, if the PCR is performed in a microchip with real-time detection using the optimized amplification mode, the result can be obtained 13–14 min after the onset of reaction.