Integrated Microfluidic Nucleic Acid Sample Preparation

Integrated Microfluidic Nucleic Acid Sample Preparation

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This dissertation presents sample preparation techniques and its integration into a microfluidic device designed to be part of a complete microscale total analysis system. A rapid prototyping technique integral to the development of the devices involving a PDMS/tape composite was introduced and several applications demonstrated. The rapid prototyping technique did not require any cleanroom facilities and made it possible to create a membrane-embedded system with ease. Based on this technique, a microfluidic genomic DNA extraction system was built that incorporated a nanoporous aluminum oxide membrane. The system was tested with lysed human blood, and rapid and inexpensive gDNA extraction was demonstrated with a high DNA yield under appropriate buffer and flow conditions. Furthermore, the extracted samples using this system were amplified without any inhibition. As an extension of this research, a high-throughput DNA extraction system was fabricated using a patterned aluminum oxide membrane. The membrane was patterned with 48 microwells shaped using SU-8 lithography and integrated with a microfluidic system. gDNA samples were flowed through the patterned membrane and the gDNA was collected on top of microwells. To track the gDNA, SYBR green was used and imaged to evaluate the variation of concentration of gDNA in each well. This high-throughput system was shown to be a useful approach for DNA extraction followed by sample distribution for use in a multiplexed assay. Using a similar concept, a semiautomatic microfluidic RNA extraction system was developed with an integrated silica membrane unit. The RNA extraction system consists of a sample container, microvalve, silica membrane unit, pneumatic micropump, and a pump controller. In this work, increasing contact times between the nucleic acid and the silica structure was found to increase the binding of RNA by up to 50% aˆ¼70%. This extraction system was integrated with a control system that reduced the user interface time and minimized the total assay time. By embedding the pneumatic micropump, it was possible for the extracted nucleic acid to be transferred to an analysis system without any addition components. This sample preparation approach for nucleic acids should be compatible with most microfluidic biosensors and other downstream assays.PAPER www.rsc.org/loc | Lab on a Chip Quantitative and qualitative analysis of a microfluidic DNA extraction system using a nanoporous A\Ox ... pore sizes and measuring the amount of hgDNA deposited on the membrane while also varying salt concentration in the solution. ... The ability to process raw samples and subsequently perform the required analytical operations on-chip is key to the eventualanbsp;...


Title:Integrated Microfluidic Nucleic Acid Sample Preparation
Author:
Publisher:ProQuest - 2009
ISBN-13:

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