Microchip large volume polymerase chain reaction (PCR) with integrated real-time capillary electrophoresis detection

摘要

A microfluidic device is provided with appropriate integrated structures to carry out large volume PCR and end-point or real-time capillary electrophoresis detection. The microfluidic device includes a substrate having a chamber for amplification of a volume of nucleic acid; wells disposed on the substrate; flow channels connecting the wells and the chamber in the substrate to allow for flow of solution through the chamber; and one or more separation channels provided in the substrate and connected to the chamber for separating and detecting a fraction of the amplified nucleic acid in the chamber. The chamber, the flow channels, and the one or more separation channels are configured such that the hydrodynamic flow resistance of the chambers and the flow channels combined is at least 10³ times smaller than the hydrodynamic flow resistance in the one or more separation channels. The microfluidic device can achieve a very high sensitivity in detection while being highly cost effective.