| Summary: | 碩士 === 中原大學 === 醫學工程研究所 === 94 === Gel electrophoresis (GE) is a technology that separates macromolecules such as DNA segments or proteins. These macromolecules possess electric charges which can be attracted by an applied electric field. Consequenctly they move through pores within a piece of porous gel. The mobility depends on the strength of applied electric field, electric charge, size, and shape of the molecules, pH value and temperature of buffer, porous size of the gel. The GE technology plays an important role in biotechnology. Any improvement to the GE technology can be significantly impact to the development of biotech.
Usually the mobility and the separation resolution of the macromolecules are considered to be performance factors of a GE system. In this course, a few approaches were used to enhance the system performance. First, higher voltages were applied on the GE bath with a thermal control system which is mainly to sink the heat of the buffer. The apparatus was then used in conjunction with an ultrasonic system. Voltages applied in the studies included 100, 150, and 200 volts. To study the thermal effects, the temperature in the GE bath was controlled to be as steady as possible that ranges between 20 and 50 ℃ with 5 ℃ increment. When the ultrasonic system was applied, the output power was set on 150 watts. The major materials used in this course are agarous and 2-Log DNA Ladder marker.
The study results show that by controlling the GE bath temperature at 20, 25, or 30 ℃ when the applied voltages increased from 100 to 200 volts, the mobility speeded up 131% on average. This is considered as a significant enhancement. By fixing an applied voltage on 100, 150, or 200 volts, when the GE bath temperature increased, the mobility increased as well. For the GE bath temperatures between 20 ℃and 50 ℃ that used in the studies, we notice that there is 2.67% increase of mobility on average. In the ultrasound studies, the factors of GE bath temperature (30 ℃~ 50 ℃) and and applied voltage (100、150 or 200 Volts) were also considered. The results show that on average the mobility increases 10.5%; however, the variation is rather large that the highest mobility is 14.5% and the lowest mobility is 2.1%. It must be noted that the DNA band resolution is independent on the GE bath temperature, the applied voltage and the ultrasounic action.
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