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Hayes CJ, Dalton TM. Microfluidic droplet-based PCR instrumentation for high-throughput gene expression profiling and biomarker discovery. Biomol Detect Quantif. 2015;4:22-32doi: 10.1016/j.bdq.2015.04.003.
Hayes, C. J., & Dalton, T. M. (2015). Microfluidic droplet-based PCR instrumentation for high-throughput gene expression profiling and biomarker discovery. Biomolecular detection and quantification, 422-32. https://doi.org/10.1016/j.bdq.2015.04.003
Hayes, Christopher J, and Dalton, Tara M. "Microfluidic droplet-based PCR instrumentation for high-throughput gene expression profiling and biomarker discovery." Biomolecular detection and quantification vol. 4 (2015): 22-32. doi: https://doi.org/10.1016/j.bdq.2015.04.003
Hayes CJ, Dalton TM. Microfluidic droplet-based PCR instrumentation for high-throughput gene expression profiling and biomarker discovery. Biomol Detect Quantif. 2015 Jun 06;4:22-32. doi: 10.1016/j.bdq.2015.04.003. eCollection 2015 Jun. PMID: 27077035; PMCID: PMC4822205.
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Biomol Detect Quantif. 2015 Jun 06;4:22-32. doi: 10.1016/j.bdq.2015.04.003. eCollection 2015 Jun.

Microfluidic droplet-based PCR instrumentation for high-throughput gene expression profiling and biomarker discovery.

Biomolecular detection and quantification

Christopher J Hayes, Tara M Dalton

Affiliations

  1. Stokes Institute, Department of Mechanical, Aeronautical and Biomedical Engineering, University of Limerick, Limerick, Ireland; Department of Life Sciences, University of Limerick, Limerick, Ireland.
  2. Stokes Institute, Department of Mechanical, Aeronautical and Biomedical Engineering, University of Limerick, Limerick, Ireland.

PMID: 27077035 PMCID: PMC4822205 DOI: 10.1016/j.bdq.2015.04.003

Abstract

PCR is a common and often indispensable technique used in medical and biological research labs for a variety of applications. Real-time quantitative PCR (RT-qPCR) has become a definitive technique for quantitating differences in gene expression levels between samples. Yet, in spite of this importance, reliable methods to quantitate nucleic acid amounts in a higher throughput remain elusive. In the following paper, a unique design to quantify gene expression levels at the nanoscale in a continuous flow system is presented. Fully automated, high-throughput, low volume amplification of deoxynucleotides (DNA) in a droplet based microfluidic system is described. Unlike some conventional qPCR instrumentation that use integrated fluidic circuits or plate arrays, the instrument performs qPCR in a continuous, micro-droplet flowing process with droplet generation, distinctive reagent mixing, thermal cycling and optical detection platforms all combined on one complete instrument. Detailed experimental profiling of reactions of less than 300 nl total volume is achieved using the platform demonstrating the dynamic range to be 4 order logs and consistent instrument sensitivity. Furthermore, reduced pipetting steps by as much as 90% and a unique degree of hands-free automation makes the analytical possibilities for this instrumentation far reaching. In conclusion, a discussion of the first demonstrations of this approach to perform novel, continuous high-throughput biological screens is presented. The results generated from the instrument, when compared with commercial instrumentation, demonstrate the instrument reliability and robustness to carry out further studies of clinical significance with added throughput and economic benefits.

Keywords: Biomarkers; Droplet; Gene expression; Instrumentation; Microfluidic; Real-time PCR

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