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 µL) 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 µL; 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.
A n electrowetting-on-dielectric (EWOD) microfluidic system of coplanar electrode type was designed for parallel DNA ligation of an ultra-micro volume, which was applied to droplet manipulation containing DNA, a ligation enzyme and a multi-salt reaction buffer. For environmental reasons, we applied a concept of a watereoil coreeshell droplet; such droplets of volume w0.3 mL were generated from reservoirs between one plate with coplanar electrodes and another plate with a hydrophobic surface free of electrodes. The experimental results show that the best mixing sequence for the DNA ligation involves mixing first the insert DNA and vector DNA, and then adding the ligase solution. The waiting time was less than 5 min. In one cloning test, the total usage of reagents in an ultramicro-volume EWOD chip was 2.1 mL with no wasted volume, for comparison with 85% waste according to the standard protocol was 15 mL. The results showed also that four parallel DNA ligations were accomplished without damage to a chip and without biomaterial annulment. ( JALA 2010;15:210-5)
The aim of this research is to develop a light ac tuated droplet manipulation system. This research will utilize ultraviolet to activate oxidation-reduction mechanism of nano-Ti02 photocatalyst coating on the base material. The water-affinity of the base material will changed due to the decrease free-energy of the material surface. The contact angle between the liquid droplet and base material will also be changed to facilitate the manipulation purpose. In this study.the system that was developed includes which ultraviolet masking device, moving platform, main frame structure etc.The software was developed by using Microsoft Visual Basic 6.0. Upon completion of hardware and software integration, experiments were carried out to verified system functionalities.This study developed a non-continuous optD-wetting droplet manipulation technique. By exposure to 6.8mW UV light, the surface tension of coating nano-Ti02 (Anatase type. 3.5 0/ .. , PH 1.5 • and with average diameter of 69 nm.)will be changed for the sake of changing hydrophilic. The change of surface tension will be applied to drag the droplet. The moving rate of dragging was 3.33mmlsec. This technique will overcome the shortcoming of opto-electrowetting technique. The program ming function of this technique will also be applied to the clinic test in the future. Index Terms -TiO} • microjluidics • manipulation of liquid droplet • opto-wetting 1. INTRODUCTION The droplet manipulation is the core technology of bio chip. Microfl uidic devices design involves two different flow patterns -continuous flow and non-continuous flow. The continuous flow microfluidics was operated by way of continuous flow madc by pump, gate, and fluidics. The noncontinuous flow microfluidics was operated by way of manipulation of independent droplet. The trend for micro fl uidics is toward non-continuous flow type for the sake of lower cost for hardware, less specimen, and faster reaction time. What's more, the cross reaction of residual chemicals in the microfluidics can be avoided efficiently. However, the problems of surface tension must be overcome in the non continuous flow microfluidics. The literatures review pro posed the opto-electrowctting and electrochemical methods to solve the problems , and thc fonner one was the most common one. The opto-electrowetting way was applied to manipUlate the droplet by changin g the surface tension and hydrophilic of base material by way of laser and polar ar rangement. The biochip , however, becomes more and more 0-7803-8998-01051$20.00 minimal than ever. Either the smaller polar arrangement or the laser facilities was expensive and not accessible. The aim of this research is to develop a UV-light actuated droplet manipulation system. Masao (1998) manipulated the droplet under different surface tension from variable static electricity. The volume of the droplet is luL, and the fastest moving rate is 400um/sec[ 1]. Pascale (200 I) manipulated the droplet by hydrophobic powder (Silane-treated lycopodi um). The av erage diameter of powders is 2...
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