Et. Lagally et al., Monolithic integrated microfluidic DNA amplification and capillary electrophoresis analysis system, SENS ACTU-B, 63(3), 2000, pp. 138-146
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.