DNA ligation of ultramicro volume using an EWOD microfluidic system with coplanar electrodes
DNA ligation of ultramicro volume using an EWOD microfluidic system with coplanar electrodes
复制标题
DOI：
10.1088/0960-1317/18/4/045017
复制
发表时间：
2008-04-01
影响因子：
2.3
通讯作者：
Chang, Hwan-You
中科院分区：
文献类型：
--
作者：
Liu, Yi-Ju;Yao, Da-Jeng;Chang, Hwan-You
为了节省试剂，设计了一种使用共面电极型电介质润湿(EWOD)微流控系统的自动超微体积DNA连接过程。这些含有DNA、连接酶和多盐反应缓冲液的液滴使用开发的EWOD系统完成了连接。在具有共面电极的平板和无电极的疏水表面的平板之间的储液池中成功地产生了超微体积的液滴(0.3MU L)。在一次成功的克隆中，超微体积EWOD芯片中试剂的总使用量为2.1亩L；没有体积浪费，而标准方法的浪费为85%，自由盖共面EWOD芯片的浪费为80%。结果还表明，整个工艺过程不会对芯片表面造成损害，也不会导致生物材料失效。通过设计一种由柔性模式的电极路径和多个储存库组成的设计，具有共面电极的EWOD数字微流控系统将被改进为一个高效的并行DNA克隆系统，用于人工文库或表达文库的构建。
An automatic ultramicro-volume DNA ligation process using a coplanar electrode type of electrowetting-on-dielectric (EWOD) microfluidic system was designed for economy of reagent. The droplets, containing DNA, a ligation enzyme and a multi-salt reaction buffer, served to complete the ligation using a developed EWOD system. Droplets of ultramicro volume (0.3 mu L) were successfully generated from reservoirs between one plate with coplanar electrodes and another plate with a hydrophobic surface free of electrodes. In one successful cloning, the total usage of reagents in an ultramicro-volume EWOD chip was 2.1 mu L; no volume was wasted, in comparison with 85% waste with the standard protocol and 80% waste with a free-cover coplanar EWOD chip. The results also show that the entire process was accomplished without damage to the chip surface and without biomaterial annulment. With a design consisting of electrode pathways in a flexible pattern and multiplex reservoirs, an EWOD digital microfluidic system with coplanar electrodes would be improved as an efficient parallel DNA-cloning system in the construction of an artificial library or expression library.