Microfluidic patterning of miniaturized DNA arrays on plastic substrates

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DOIResolve DOI: http://doi.org/10.1021/am900285g
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Journal titleACS Applied Materials and Interfaces
Pages13871395; # of pages: 9
SubjectDNA patterning; DNA arrays; plastic surfaces; plastic substrates; microcapillary system; microspotting; poly(dimethylsiloxane); copolymers; fluidic patterning
AbstractThis paper describes the patterning of DNA arrays on plastic surfaces using an elastomeric, two dimensional (2D) microcapillary system (μCS). Fluidic structures were realized through hot embossing lithography using Versaflex® CL30. Like elastomers based on poly(dimethylsiloxane) (PDMS), this thermoplastic block co-polymer is able to seal a surface in a reversible manner, making it possible to confine DNA probes with a level of control that is unparalleled using standard microspotting techniques. We focus on μCSs that support arrays comprising up to 48 spots each being 45 μm in diameter. Substrates were fabricated from two hard termoplastic materials - poly(methylmethacrylate) (PMMA) and a polycyclic olefin (e.g., Zeonor® 1060R) - which were both activated with N-hydroxysuccinimide (NHS) ester to mediate covalent attachment of DNA molecules. The approach was exemplified by using 0.25 to 32 μM solutions of amino-modified oligonucleotides labeled with either Cy3 or Cy5 fluorescent dye in phosphate buffered saline (PBS), allowing for a direct and sensitive characterization of the printed arrays. Solutions were incubated for durations of 1 to >48 h at22, 30 and 40℃ to probe the conditions for obtaining uniform spots of high fluorescence intensity. The length (l) and depth (d) of microfluidic supply channels were both important with respect to depletion as well as evaporation of the solvent. While selective activation of the substrate proved helpful to limit unproductive loss of oligonucleotides along trajectories, incubation of solution in a humid environment was necessary to prevent uncontrolled drying of liquid, keeping the immobilization process intact over extended periods of time. When combined, these strategies effectively promoted the formation of high-quality DNA arrays, making it possible to arrange multiple probes in parallel with a high degree of uniformity. Moreover, we show that resultant arrays are compatible with standard hybridization protocols, which allowed for reliable discrimination of individual stands when exposed to a specific ssDNA target molecule.
Publication date
PublisherAmerican Chemical Society
Copyright noticeMaterial in this document is covered by the provisions of the Copyright Act, by Canadian laws, policies, regulations and international agreements. Such provisions serve to identify the information source and, in specific instances, to prohibit reproduction of materials without written permission.
AffiliationNRC Industrial Materials Institute (IMI-IMI); NRC Steacie Institute for Molecular Sciences (SIMS-ISSM); National Research Council Canada
Access conditionavailable
Peer reviewedYes
NRC number51093
NPARC number10944274
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Record identifiere9d6fff8-cb4c-4e5d-a28a-adad8cca7068
Record created2009-10-03
Record modified2016-05-09
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