Temperature and time-resolved total internal reflectance fluorescence analysis of reusable DNA hydrogel chips.
Temperature and time-resolved total internal reflectance fluorescence analysis of reusable DNA hydrogel chips.
复制标题
可重复使用 DNA 水凝胶芯片的温度和时间分辨全内反射荧光分析
DOI：
10.1021/ac1008578
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发表时间：
2010
影响因子：
7.4
通讯作者：
Rühe J.
中科院分区：
文献类型：
--
作者：
Neumann T;Bonham A. J;Dame G;Bertold B;Brandstetter T.;Rühe J.
全内反射荧光（TIRF）与共价结合在PMMA基底上的水凝胶-DNA微滴微阵列相结合，为评估DNA杂交和快速生物芯片开发提供了一个可重复使用的灵敏平台。水凝胶微阵列，其中包含共价结合的DNA探针，是通过一个简单的印刷和光交联过程。TIRF测量的阵列显示强大的可重复使用性，显示线性灵敏度低至5 fmol的荧光标记的靶DNA，是敏感的单碱基对错配。此外，通过PCR扩增杂交的研究显示了询问较大DNA的能力。我们最终证明了一个有效的，可重复的，低成本的DNA杂交研究平台，可用于快速高通量诊断以及生物芯片的开发。
Total internal reflection fluorescence (TIRF) coupled with hydrogel−DNA droplet microarrays covalently bound on PMMA substrates presents a reusable, sensitive platform for evaluating DNA hybridization and for rapid biochip development. Hydrogel microarrays, which contain covalently bound DNA probes, are created via a simple printing and photocross-linking process. TIRF measurements of the arrays display robust reusability, show linear sensitivity down to 5 fmol of fluorescently labeled target DNA, and are sensitive to single basepair mismatches. Additionally, the ability to interrogate larger DNA is shown through studies with PCR amplification hybridization. We conclusively demonstrate an efficient, reproducible, low cost platform for DNA hybridization studies that could be used for fast high-throughput diagnostics as well as biochip development.