Rapid DNA detection by the real-time polimerase chain reaction

The paper concerns the optimization of conditions of Real-Time polymerase chain reaction (Real-Time PCR) taking place in a microfluidic analytical system. We have compared the methods for estimating the temperature inside the microreactors and proved that the contact method provides adequate estimates of temperature achieved inside the microfluidic chip microreactors. Optimization of the amplification regime has been carried out; it has been proved that reduction of the temperature duration at the termocycling stages allows one to decrease considerably the analysis time without losses in sensitivity and also without drastic reduction of the PCR efficiency. The paper demonstrates that the time necessary to pick up 5 copies of DNA molecules is 18 minutes only.