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Published July 1, 2005 | public
Journal Article

Electrokinetically Based Approach for Single-Nucleotide Polymorphism Discrimination Using a Microfluidic Device

Abstract

In this work, we describe and implement an electrokinetic approach for single-nucleotide polymorphism (SNP) discrimination using a PDMS/glass-based microfluidic chip. The technique takes advantage of precise control of the coupled thermal (Joule heating), shear (electroosmosis), and electrical (electrophoresis) energies present at an array of probes afforded by the application of external electrical potentials. Temperature controllers and embedded thermal devices are not required. The chips can be easily and inexpensively fabricated using standard microarray printing methods combined with soft-lithography patterned PDMS fluidics, making these systems easily adaptable to applications using higher density arrays. Extensive numerical simulations of the coupled flow and thermal properties and microscale thermometry experiments are described and used to characterize the in-channel conditions. It was found that optimal conditions for SNP detection occur at a lower temperature on-chip than for typical microarray experiments, thereby revealing the importance of the electrical and shear forces to the overall process. To demonstrate the clinical utility of the technique, the detection of single-base pair mutations in the survival motor neuron gene, associated with the childhood disease spinal muscular atrophy, is conducted.

Additional Information

© 2005 American Chemical Society. Received 7 February 2005; accepted 21 April 2005; published online 18 May 2005; published in print 1 July 2005. We are grateful to the Natural Sciences and Engineering Research Council of Canada for financial support toward this research work. D.E. also acknowledges the financial support of Glynn Williams through a scholarship. We thank Dr. Paul. A. E. Piunno in the Department of Chemistry, University of Toronto at Mississauga, and Dr. Sandeep Raha of McMaster University, for useful discussion, and for PCR amplification. We also thank Dr. Jianming Pei and Dr. Tim Westwood of the University of Toronto at Missisauga Department of Zoology for assistance with printing of DNA microarrays.

Additional details

Created:
August 19, 2023
Modified:
October 20, 2023