In situ synthesis of DNA microarray on functionalized cyclic olefin copolymer substrate.
Thermoplastic materials such as cyclic-olefin copolymers (COC) provide a versatile and cost-effective alternative to the traditional glass or silicon substrate for rapid prototyping and industrial scale fabrication of microdevices. To extend the utility of COC as an effective microarray substrate, we developed a new method that enabled for the first time in situ synthesis of DNA oligonucleotide microarrays on the COC substrate. To achieve high-quality DNA synthesis, a SiO(2) thin film array was prepatterned on the inert and hydrophobic COC surface using RF sputtering technique. The subsequent in situ DNA synthesis was confined to the surface of the prepatterned hydrophilic SiO(2) thin film features by precision delivery of the phosphoramidite chemistry using an inkjet DNA synthesizer. The in situ SiO(2)-COC DNA microarray demonstrated superior quality and stability in hybridization assays and thermal cycling reactions. Furthermore, we demonstrate that pools of high-quality mixed-oligos could be cleaved off the SiO(2)-COC microarrays and used directly for construction of DNA origami nanostructures. It is believed that this method will not only enable synthesis of high-quality and low-cost COC DNA microarrays but also provide a basis for further development of integrated microfluidics microarrays for a broad range of bioanalytical and biofabrication applications.
Saaem
Ishtiaq
I
Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
Ma
Kuo-Sheng
KS
Marchi
Alexandria N
AN
LaBean
Thomas H
TH
Tian
Jingdong
J
D003516:Cycloparaffins / Q000737:chemistry*; D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008567:Membranes, Artificial; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D011108:Polymers / Q000737:chemistry*
D003516:Cycloparaffins; D008567:Membranes, Artificial; D011108:Polymers; D004247:DNA