Monolithic integrated microfluidic DNA amplification and capillary electrophoresis analysis system

A monolithic integrated DNA analysis system comprised of microfluidic valve s and vents, PCR amplification chambers, and capillary electrophoretic sepa ration channels has been microfabricated in a glass sandwich structure. Val ves and hydrophobic vents provide controlled and sensorless sample loading into 280 nl PCR chambers; the low volume reactor and the use of thin film h eaters permit thermal cycle times as fast as 30 s. The amplified product is labeled with an intercalating fluorescent dye and directly injected into a microfabricated capillary electrophoresis channel. Analyses with this devi ce have produced and detected PCR products from reactions with as few as 20 starting DNA template copies/mu l, (five to six copies/chamber), The extra polated limit of detection based on data using 20 cycles is two copies/cham ber. The optimization of heater placement, thermal anisotropy measurements, and the optimization of thermal profiles are also discussed. (C) 2000 Else vier Science S.A. All rights reserved.