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A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons
Published on: November 14, 2018
Solid phase extraction of DNA from biological samples in a post-based, high surface area poly(methyl methacrylate)
Carmen R Reedy1, Carol W Price, Jeff Sniegowski
1Department of Chemistry, University of Virginia, McCormick Road, P.O. Box 400319, Charlottesville, Virginia 22904, USA.
Lab on a Chip
|March 8, 2011
Summary
This study developed poly(methyl methacrylate) microfluidic devices for DNA purification using chitosan-functionalized posts. These devices successfully purified human genomic DNA from whole blood for PCR amplification.
Area of Science:
- Biomaterials Science
- Microfluidics
- Molecular Biology
Background:
- Poly(methyl methacrylate) (PMMA) offers advantages for microfluidic device fabrication due to reproducible high surface area (SA) structures via X-ray lithography.
- Challenges include post fragility during bonding and effective surface functionalization for DNA capture.
Purpose of the Study:
- To develop and evaluate PMMA-based microfluidic DNA purification devices with functionalized microfabricated posts.
- To optimize post geometry for high surface area and low fluidic resistance.
- To demonstrate efficient DNA purification from biological samples for downstream applications.
Main Methods:
- Covalent functionalization of PMMA posts with chitosan for pH-dependent DNA binding.
- Development of bonding techniques to preserve post integrity in closed microfluidic systems.
- Exploration of various post geometries to balance surface area and fluidic resistance.
Main Results:
- Successful covalent functionalization of PMMA posts with chitosan.
- Demonstration of pH-dependent DNA binding and elution.
- Identification of post designs balancing surface area and fluidic resistance.
- Recovery of prepurified human genomic DNA (hgDNA) from whole blood.
- Confirmation of purified hgDNA's suitability for PCR amplification.
Conclusions:
- PMMA microfluidic devices with chitosan-functionalized posts provide an effective platform for DNA purification.
- The developed methods overcome challenges in PMMA microfluidic fabrication and functionalization.
- The devices show promise for real-world applications, such as purifying DNA from blood for genetic analysis.
