Monolithic integrated PCR reactor-CE system for DNA amplification and analysis to the single molecule limit

A fully integrated genomic analysis microsystem including heaters, temperature sensors, and polymerase chain reaction (PCR) chambers directly connected to capillary electrophoresis (CE) separation channels has been developed. The device utilizes pneumatic valves and hydrophobic vents for positive and sensorless sample loading and immobilization within 200 nL PCR chambers. Use of microfabrication techniques for the creation of resistive heating and temperature-sensing elements has improved the thermal response and accuracy of the system beyond previous limits. Amplified PCR products are labeled on-column with a fluorescent intercalating dye and injected into a gel-filled microchannel for electrophoretic analysis. Successful sex determination by performing multiplex PCR from human genomic DNA is demonstrated in under 15 minutes.