Onoshima Daisuke, Baba Yoshinobu
Institute of Innovation for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
ImPACT Research Center for Advanced Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Methods Mol Biol. 2017;1547:105-111. doi: 10.1007/978-1-4939-6734-6_8.
The method described here enables the automatic stretching and patterning of single DNA molecules onto a solid surface. It does not require chemical modification of the DNA or surface modification of the substrate. To detect a signal variation caused by sequence-specific dye binding or partial melting, it is crucial that the DNA molecules are arrayed in a parallel direction inside the narrow microscopic field. The method uses zigzag-shaped microgrooves in a densely-arranged molecular patterning apparatus in a microfluidic channel. By syringing through the microchannel, over 1500 DNA molecules can be arrayed simultaneously in the microgrooves. It will therefore serve as a template preparation for DNA molecular diagnosis by high-resolution imaging.
这里所描述的方法能够将单个DNA分子自动拉伸并排列到固体表面上。它不需要对DNA进行化学修饰,也不需要对底物进行表面修饰。为了检测由序列特异性染料结合或部分解链引起的信号变化,至关重要的是DNA分子要在狭窄的微观视野内沿平行方向排列。该方法在微流控通道中密集排列的分子图案化装置中使用锯齿形微槽。通过向微通道中注射,1500多个DNA分子可以同时排列在微槽中。因此,它将作为通过高分辨率成像进行DNA分子诊断的模板制备方法。