Combining Electro-Osmotic Flow and FTA Paper for DNA Analysis on Microfluidic Devices.

Micromachines (Basel)

Faculty of Science and Engineering, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK.

Published: July 2016

FTA paper can be used to protect a variety of biological samples prior to analysis, facilitating ease-of-transport to laboratories or long-term archive storage. The use of FTA paper as a solid phase eradicates the need to elute the nucleic acids from the matrix prior to DNA amplification, enabling both DNA purification and polymerase chain reaction (PCR)-based DNA amplification to be performed in a single chamber on the microfluidic device. A disc of FTA paper, containing a biological sample, was placed within the microfluidic device on top of wax-encapsulated DNA amplification reagents. The disc containing the biological sample was then cleaned up using Tris-EDTA (TE) buffer, which was passed over the disc, via electro-osmotic flow, in order to remove any potential inhibitors of downstream processes. DNA amplification was successfully performed (from buccal cells, whole blood and semen) using a Peltier thermal cycling system, whereupon the stored PCR reagents were released during the initial denaturing step due to the wax barrier melting between the FTA disc and PCR reagents. Such a system offers advantages in terms of a simple sample introduction interface and the ability to process archived samples in an integrated microfluidic environment with minimal risk of contamination.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6190317PMC
http://dx.doi.org/10.3390/mi7070119DOI Listing

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