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Polymeric microfluidic devices for DNA analysis
Author
Sun, Yi
Supervisor
Kwok, Yien Chien
Abstract
DNA amplification and separation are two vital processes in DNA analysis. Towards the ultimate goal of developing a fully integrated microfluidic device for “sample-to-answer” analysis, in this project, we design and fabricate polymerase chain reaction (PCR) microchips and capillary electrophoretic (CE) microchips for fast amplification and precise separation of DNA samples. The microchips are fabricated in poly(methyl methacrylate) (PMMA) substrates. A novel thermal bonding process with high temperature and low bonding pressure is developed for assembling PMMA-sheets. A hybrid PMMA-Polycarbonate (PC) continuous flow PCR microchip is fabricated and has been proven to have better heat tolerance. Novel ferrofluid driven single-channel and multi-channel PCR microchips are also developed. Amplification of 500 bp lambda DNA fragments, forensic DNA samples and genetic modified organism (GMO) samples are successfully performed. A micro CE chip is also fabricated and has been tested by performing gel electrophoretic separation of 11-fragment X174-Hae III dsDNA ladders and peak tailing problem has been encountered. To tackle peak tailing, a mathematical model is developed to identify the primary cause. Based on the model, the microfluidic CE device is modified and the feasibility for DNA separation is investigated by performing rapid and high-resolution electrophoretic separations of nine allelic miniD16 ladder. To improve separation efficiency, a novel capillary bundle microchip is developed to perform CE in extremely narrow channels by using photonic crystal fiber (PCF) as separation column. The capillary bundle offers rapid heat dissipation, which enables the use of ultra high electric field for separation.
Date Issued
2010
Call Number
QP624 Sun
Date Submitted
2010