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

Abstract

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.

Valve‐Enabled Sample Preparation and RNA Amplification in a Coffee Mug for Zika Virus Detection

“…Examples of these products are FTAcard, [16][17][18][19][20][21][22] chitosancoated paper, [23,24] glass-fiber paper, [25][26][27][28] and polyethersulfone filter paper. Examples of these products are FTAcard, [16][17][18][19][20][21][22] chitosancoated paper, [23,24] glass-fiber paper, [25][26][27][28] and polyethersulfone filter paper.…”

Towards Multiplex Molecular Diagnosis—A Review of Microfluidic Genomics Technologies

“…Several groups have developed workarounds. Liu et al [ 79 ] and Wimbles et al [ 80 ] also loaded amplification reagents onto FTA filters. Liu demonstrated extraction and isothermal amplification of viral nucleic acid, while Wimbles incorporated electro-osmotic flow (EOF) to transport washing buffers to the membrane ( Figure 5 ).…”

“… ( a ) Microfluidic device used to perform integrated DNA purification and amplification experiments; ( b ) schematic top view of the channels and central chamber; ( c ) schematic cross section view of FTA ® paper discs placed on top of a layer of wax-encapsulated PCR. Reproduced with permission from [ 80 ]. …”

Application of Microfluidic Methodology for the Analysis of DNA

“…In the often neglected area of sample introduction, the paper by Wimbles et al [ 3 ] describes the extraction of DNA directly from FTA ® paper used to collect, lyse and preserve a range of sample types of forensic interest. The samples are able to be added directly to a chamber in the glass microfluidic device on top of pre-loaded PCR reagents which are protected by a waxy layer.…”