Hydrogel drop microchips with immobilized DNA: properties and methods for large-scale production

Although gel-based microchips offer significant advantages over two-dimensional arrays, their use has been impeded by the lack of an efficient manufacturing procedure. Here we describe two simple, fast, and reproducible methods of fabrication of DNA gel drop microchips. In the first, copolymerizatio...

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Bibliographic Details
Published in:Analytical biochemistry 2004-02, Vol.325 (1), p.92-106
Main Authors: Rubina, A.Yu, Pan’kov, S.V, Dementieva, E.I, Pen’kov, D.N, Butygin, A.V, Vasiliskov, V.A, Chudinov, A.V, Mikheikin, A.L, Mikhailovich, V.M, Mirzabekov, A.D
Format: Article
Language:English
Subjects:
Acrylamides - chemistry
Copolymerization
DNA - chemistry
DNA microarrays
Gel microchips
Hybridization, Genetic
hydrogel microchips
Hydrogel, Polyethylene Glycol Dimethacrylate - chemical synthesis
hydrogels
IMAGE chips
Microchip manufacturing
microchips
Oligonucleotide Array Sequence Analysis - instrumentation
Oligonucleotide Array Sequence Analysis - methods
Oligonucleotides - chemical synthesis
Oligonucleotides - chemistry
Photochemistry
Polymers - chemistry
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Fluorescence
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Summary:Although gel-based microchips offer significant advantages over two-dimensional arrays, their use has been impeded by the lack of an efficient manufacturing procedure. Here we describe two simple, fast, and reproducible methods of fabrication of DNA gel drop microchips. In the first, copolymerization method, unsaturated groups are chemically attached to immobilized molecules, which are then mixed with gel-forming monomers. In the second, simpler polymerization-mediated immobilization method, aminated DNA without prior modification is added to a polymerization mixture. Droplets of polymerization mixtures are spotted by a robot onto glass slides and the slides are illuminated with UV light to induce copolymerization of DNA with gel-forming monomers. This results in immobilization of DNA within the whole volume of semispherical gel drops. The first method can be better controlled while the second one is less expensive, faster, and better suited to large-scale production. The microchips manufactured by both methods are similar in properties. Gel elements of the chip are porous enough to allow penetration of DNA up to 500 nucleotides long and its hybridization with immobilized oligonucleotides. As shown with confocal microscope studies, DNA is hybridized uniformly in the whole volume of gel drops. The gels are mechanically and thermally stable and withstand 20 subsequent hybridizations or 30–40 PCR cycles without decrease in hybridization signal. A method for quality control of the chips by staining with fluorescence dye is proposed. Applications of hydrogel microchips in research and clinical diagnostics are summarized.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2003.10.010