膠體電泳設置改良對DNA分離效益之探討

膠體電泳是一項分離DNA片段和蛋白質等生物分子的技術。其原理是藉電場強度作用於具電荷之生物分子上，引動分子於膠體孔隙運動；運動速率大致與使用的電場強度、分子電荷量多寡、分子大小、分子形狀、緩衝溶液酸鹼值、溫度和膠體孔洞大小相關。在生物科技領域裡，膠體電泳扮演著重要角色，任何電泳技術的提升，對生物科技的發展將有深遠地影響。一般而言，泳動速率和分子聚集帶解析度是電泳系統分離生物分子二項重要的參數。本研究以高電壓搭配溫控系統應用在電泳系統中，有效的降低緩衝液的熱度而提高電泳速率。上述裝置同時結合超音波系統，嘗試找出提升速率的參數。實驗中所使用的參數值如下：包括的電壓值有100、150和200 V；溫度值則從20 ℃開始以每5 ℃增加到50 ℃；超音波功率則是以150 W應用在實驗之中。所使用的主要材料是洋菜膠和2-Log DNA Ladder (0.1-10.0 Kb)標記。研究結果顯示，電泳槽溫度控制在20、25或30 ℃，且電壓範圍由100 V提高至200 V時，各組泳動速率比值有明顯的增加，各比值也相當接近，平均提高1.31倍。電泳槽電壓固定為100、150或200 V，溫度範圍由20 ℃提高至50 ℃時；這樣條件下所獲得的實驗結果顯示，每升高1 ℃，所對應之泳動速率平均提高約2.67%。至於超音波對電泳的作用實驗，也是以控溫和定壓的條件進行，從30 ℃增加到50 ℃及100、150和200 V；結果顯示，有超音波作用比起無超音波作用所獲得的平均泳動率提高10.5%，但最大為14.5%，最小為2.1%，顯示有相當的變異。值得注意的是，DNA在電泳系統作用下的分子聚集帶解析度與溫度、電壓和有無超音波系統的作用並不相關。

關鍵字

溫控系統；焦耳熱；電泳；超音波

並列摘要

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.

並列關鍵字

Joule heat ； Temperature control system ； Ultrasound ； Electrophoresis

參考文獻

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膠體電泳設置改良對DNA分離效益之探討

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