Functional Integration of PCR Amplification and Capillary Electrophoresis in a Microfabricated DNA Analysis Device

Abstract

Microfabricated silicon PCR reactors and glass capillary electrophoresis (CE) chips have been successfully coupled to form an integrated DNA analysis system. This construct combines the rapid thermal cycling capabilities of microfabricated PCR devices (10 °C/s heating, 2.5 °C/s cooling) with the high-speed (Salmonella DNA was performed in under 45 min, demonstrating that challenging amplifications of diagnostically interesting targets can also be performed. Real-time monitoring of PCR target amplification in these integrated PCR−CE devices is also feasible. Amplification of the β-globin target as a function of cycle number was directly monitored for two different reactions starting with 4 × 10⁷ and 4 × 10⁵ copies of DNA template. This work establishes the feasibility of performing high-speed DNA analyses in microfabricated integrated fluidic systems.