Microfluidic transfer of liquid interface for parallel stretching and stamping of terminal-unmodified single DNA molecules in zigzag-shaped microgrooves
Microfluidic transfer of liquid interface for parallel stretching and stamping of terminal-unmodified single DNA molecules in zigzag-shaped microgrooves
复制标题
液体界面的微流体转移，用于在锯齿形微槽中平行拉伸和冲压末端未修饰的单个 DNA 分子
DOI：
10.1039/c4lc00990h
复制
发表时间：
2014
期刊：
影响因子：
6.1
通讯作者：
and Y. Baba
中科院分区：
文献类型：
--
作者：
H. Yasaki;D. Onoshima;T. Yasui;H. Yukawa;N. Kaji;and Y. Baba
DNA的分子拉伸是在单分子水平上对DNA的碱基序列和表观基因组变化进行光学探索的不可或缺的工具。在拉伸末端未修饰的DNA分子在固体基底上彼此平行时，通过去湿过程的后退弯月面组装和毛细管力是非常有用的。这些可以通过将基底从DNA溶液中拉出或在一对基底之间滑动DNA溶液的液滴来实现。然而，目前使用的方法不允许控制液体界面运动和单分子DNA定位。在这里，我们展示了一种微流体装置，通过微槽拉伸DNA分子。该装置可以在微槽的顶点捕获单个DNA分子，微槽被设计为平行之字形线。不同的锯齿形图案的深度，大小和形状进行了研究，以评估DNA在表面上的吸附的可能性。液体界面的微流体转移同时拉伸超过1500个DNA分子。拉伸的DNA分子可以被压印到硅烷化的表面上。因此，该设备将作为高分辨率DNA成像研究的模板制备。
The molecular stretching of DNA is an indispensable tool for the optical exploration of base sequences and epigenomic changes of DNA at a single molecule level. In stretching terminal-unmodified DNA molecules parallel to each other on solid substrate, the receding meniscus assembly and capillary force through the dewetting process are quite useful. These can be achieved by pulling the substrate out of the DNA solution or sliding a droplet of DNA solution between a pair of substrates. However, currently used methods do not allow control over liquid interface motion and single-molecule DNA positioning. Here, we show a microfluidic device for stretching DNA molecules by syringing through microgrooves. The device can trap single DNA molecules at vertices of the microgrooves, which were designed as parallel zigzag lines. Different zigzag pattern depths, sizes, and shapes were studied to evaluate the adsorption possibility of DNA on the surface. The microfluidic transfer of the liquid interface stretched over 1500 DNA molecules simultaneously. The stretched DNA molecules could be stamped to a silanized surface. The device will therefore serve as a template preparation for high-resolution DNA imaging studies.