On-chip Device for Isothermal, Chemical Cycling Polymerase Chain Reaction

We demonstrate a novel chemical cycling polymerase chain reaction (ccPCR) technique for DNA amplification in a fully working device where temperature is held constant in space and time. We demonstrate successful ccPCR amplification while simultaneously focusing products via isotachophoresis (ITP) for identification of the environmental bacteria E. Coli. We electrophoretically drive the DNA sample with ITP through a series of high denaturant concentration zones. The denaturant is neutral so the DNA experiences alternatively low and high concentrations. This effectively replaces the thermal cycling of classical PCR (Figure 1). We performed ccPCR with end-point detection and real-time fluorescence monitoring.