Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA.
ACS Appl Mater Interfaces. 2010 Feb;2(2):491-7. doi: 10.1021/am900884b.
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.
热塑性材料,如环状烯烃共聚物(COC),为快速原型制作和工业规模制造微器件提供了一种通用且具有成本效益的替代传统玻璃或硅基板的方法。为了扩展 COC 作为有效微阵列基板的用途,我们开发了一种新方法,首次能够在 COC 基板上原位合成 DNA 寡核苷酸微阵列。为了实现高质量的 DNA 合成,使用射频溅射技术在惰性和疏水性 COC 表面预先图案化 SiO2 薄膜阵列。随后,通过喷墨 DNA 合成仪精确输送亚磷酰胺化学物质,将原位 DNA 合成限制在预图案化的亲水性 SiO2 薄膜特征的表面上。原位 SiO2-COC DNA 微阵列在杂交实验和热循环反应中表现出优异的质量和稳定性。此外,我们证明可以从 SiO2-COC 微阵列上切割出高质量的混合寡核苷酸,并直接用于构建 DNA 折纸纳米结构。相信这种方法不仅能够合成高质量、低成本的 COC DNA 微阵列,还为进一步开发用于广泛的生物分析和生物制造应用的集成微流控微阵列提供了基础。