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Rapid multiplex DNA amplification on an inexpensive microdevice for human identification via short tandem repeat
Jacquelyn A DuVall1, Delphine Le Roux1, Brandon L Thompson1
1Department of Chemistry, University of Virginia, Charlottesville, VA 22904, United States.
Analytica Chimica Acta
|June 18, 2017
Summary
This study developed a rapid polymerase chain reaction (PCR) method for forensic DNA analysis. The new microdevice platform significantly speeds up short tandem repeat (STR) profiling, enabling faster results for forensic investigations.
Area of Science:
- Forensic Science
- Molecular Biology
- Microfluidics
Background:
- Forensic DNA analysis involves multiple time-consuming steps, with polymerase chain reaction (PCR) amplification being a significant bottleneck.
- Optimizing PCR is crucial for accelerating the entire forensic DNA analysis workflow.
Purpose of the Study:
- To develop a rapid, multiplexed PCR method for forensic short tandem repeat (STR) analysis.
- To demonstrate the feasibility of integrating this rapid PCR into a microdevice platform for forensic applications.
Main Methods:
- Designed primers for 10 human genomic loci with short amplicons (<350 bp) for microchip electrophoresis compatibility.
- Optimized primer concentrations for microdevice amplification to achieve balanced STR profiles.
- Investigated rapid, multiplexed PCR on various substrates (polyethylene terephthalate) and adhesives, utilizing a custom fluid control and thermocycling system.
Main Results:
- Achieved rapid STR-based multiplexed PCR amplification in 15 minutes for a 10-plex and 10 minutes for a 6-plex (CODIS loci + Amelogenin) on a microdevice.
- Demonstrated successful amplification on diverse substrates and with different adhesives.
- Showcased preliminary integration capabilities with upstream DNA extraction and downstream microchip electrophoresis.
Conclusions:
- The developed rapid PCR microdevice platform significantly reduces analysis time for forensic DNA profiling.
- The platform shows potential for a fully integrated, rotationally-driven microdevice for complete forensic DNA analysis, compatible with lyophilized reagents.
