Dataset Information

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.

SUBMITTER: Wimbles R

PROVIDER: S-EPMC6190317 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature

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

Wimbles Ryan R Melling Louise M LM Shaw Kirsty J KJ

Micromachines 20160714 7

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 ...[more]

PMID: 30404292

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Project description:Numerous fabrication methods have been reported for microfluidic paper-based analytical devices (μPADs) using barrier materials ranging from photoresist to wax. While these methods have been used with wide success, consistently producing small, high-resolution features using materials and methods that are compatible with solvents and surfactants remains a challenge. Two new methods are presented here for generating μPADs with well-defined, high-resolution structures compatible with solvents and surfactant-containing solutions by partially or fully fusing paper with Parafilm® followed by cutting with a CO2 laser cutter. Partial fusion leads to laminated paper (l-paper) while the complete fusion results in infused paper (i-paper). Patterned structures in l-paper were fabricated by selective removal of the paper but not the underlying Parafilm® using a benchtop CO2 laser. Under optimized conditions, a gap as small as 137 ± 22 μm could be generated. Using this approach, a miniaturized paper 384-zone plate, consisting of circular detection elements with a diameter of 1.86 mm, was fabricated in 64 × 43 mm2 area. Furthermore, these ablation-patterned substrates were confirmed to be compatible with surfactant solutions and common organic solvents (methanol, acetonitrile and dimethylformamide), which has been achieved by very few μPAD patterning techniques. Patterns in i-paper were created by completely cutting out zones of the i-paper and then fixing pre-cut paper into these openings similar to the strategy of fitting a jigsaw piece into a puzzle. Upon heating, unmodified paper was readily sealed into these openings due to partial reflow of the paraffin into the paper. This unique and simple bonding method was illustrated by two types of 3D μPADs, a push-on valve and a time-gated flow distributor, without adding adhesive layers. The free-standing jigsaw-patterned sheets showed good structural stability and solution compatibility, which provided a facile alternative method for fabricating complicated μPADs.

Dataset Information

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

Publications

Combining Electro-Osmotic Flow and FTA<sup>®</sup> Paper for DNA Analysis on Microfluidic Devices.

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