Kuo Chiung-Wen, Shiu Jau-Ye, Wei Kung Hwa, Chen Peilin
Research Center for Applied Sciences, Academia Sinica, 128, Section 2, Academia Road, Nankang, Taipei 115, Taiwan.
J Chromatogr A. 2007 Aug 31;1162(2):175-9. doi: 10.1016/j.chroma.2007.06.037. Epub 2007 Jun 27.
Gel electrophoresis and capillary gel electrophoresis are widely used for the separation of biomolecules. With increasing demand in the miniaturized devices such as lab-on-a-chip, it is necessary to integrate such a separation component into a chip format. Here, we describe a simple approach to fabricate robust three-dimensional periodic porous nanostructures inside the microchannels for the separation of DNA molecules. In our approach, the colloidal crystals were first grown inside the microchannel using evaporation assisted self-assembly process. Then the void spaces among the colloidal crystals were filled with epoxy-based negative tone photoresist (SU-8). UV radiation was used to cure the photoresist at the desired area inside the microchannel. After subsequent development and nanoparticle removal, the well-ordered nanoporous structures inside the microchannel were obtained. Our results indicated that it was possible to construct periodic porous nanostructures inside the microchannels with cavity size around 300 nm and interconnecting pores around 30 nm. The mobility of large DNA molecules with different sizes was measured as a function of the applied electric field in the nanoporous materials. It was also demonstrated that 1 kilo-base pair (kbp) DNA ladders could be separated in such an integrated system within 10 min under moderate electric field.
凝胶电泳和毛细管凝胶电泳被广泛用于生物分子的分离。随着对诸如芯片实验室等小型化设备的需求不断增加,有必要将这种分离组件集成到芯片形式中。在此,我们描述了一种在微通道内制造坚固的三维周期性多孔纳米结构以分离DNA分子的简单方法。在我们的方法中,首先使用蒸发辅助自组装过程在微通道内生长胶体晶体。然后用基于环氧树脂的负性光刻胶(SU-8)填充胶体晶体之间的空隙。使用紫外线辐射在微通道内的所需区域固化光刻胶。经过后续显影和纳米颗粒去除后,在微通道内获得了有序的纳米多孔结构。我们的结果表明,有可能在微通道内构建腔尺寸约为300 nm且互连孔约为30 nm的周期性多孔纳米结构。测量了不同大小的大DNA分子在纳米多孔材料中的迁移率与施加电场的函数关系。还证明了在中等电场下,1千碱基对(kbp)的DNA梯在这样的集成系统中可以在10分钟内分离。