Procedure nucleic acid amplification using PCR on detection with integrated microchip CE realtime

摘要

A method of amplification and detection of nucleic acids for a microfluidic system comprising: (1) providing a microfluidic system comprising: a substrate having a chamber (220) for amplifying a nucleic acid volume; a plurality of wells (240) disposed on the substrate; channels (230) flow connecting the wells (240) and the camera (220) in the substrate to allow the flow of the solution through the chamber; and one or more channels (200) of separation provided on the substrate connecting the camera and other wells to separate and detect a fraction of the amplified nucleic acid in the camera, the camera (220) being configured channels (230) flow and one or more channels (200) apart so that the hydrodynamic flow resistance in the chambers and flow channels combined is at least 103 times smaller than the hydrodynamic flow resistance in one or more separation channels; (2) providing a nucleic acid solution in the chamber (220); (3) amplifying the nucleic acid disposed in the chamber (220); (4) enter from the chamber (220) a fraction of the amplified nucleic acid in one or more channels (200) apart; (5) separating the fraction of the amplified nucleic acid through one or more channels (200) apart; and (6) detecting the separated nucleic acid, wherein steps (3) to (6) are repeated one or more times, or step (3) is repeated one or more times and then steps (4) to ( 6) are executed once.