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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37103
標題: | 以多端點膠體電泳技術進行生物分子自動純化之研究 A Study on the Automated Purification of Biomolecules by Multiterminal Gel Electrophoresis |
作者: | Chun-Feng Kao 高群豐 |
指導教授: | 陳林祈(Lin-Chi Chen) |
關鍵字: | 十字形電泳通道生物分子回收,多端點電泳裝置,自動DNA分離純化,膠體電泳, automated separation,biomolecule recovery,cross channels,gel electrophoresis,multiterminal biochip, |
出版年 : | 2008 |
學位: | 碩士 |
摘要: | 近年來,由於生物領域的蓬勃發展與醫療意識的抬頭,使生物性檢測裝置快速地發展開來。有鑑於自動化技術在生物樣本純化上擁有有相當大的成長空間,本研究嘗試以光機電整合技術,發展出一套快速自動化生物分子純化電泳系統,進而應用在自動化篩選、純化生物性樣本或是DNA定序等方向,並幫助研究人員以自動化技術進行各類核酸實驗,節省以往所需耗費的人力與時間。
基於上述動機,本研究在系統開發上先以石英管柱膠體電泳進行生物性樣本的即時偵測。實驗結果證明此系統能成功將DNA ladder分離,但在後續的純化回收上有其困難之處。因此,本研究另行發展出一套多端點膠體電泳裝置,其元件以壓克力為基材,經由機械加工,將電泳通道佈於其上,並搭配光電二極體和綠光LED或UV光源,形成一套三明治結構的實驗模組。同時為了將自動化運用在電泳系統上,在後端部分接上放大電路、資料擷取卡、BJT電晶體和繼電器,加上LabVIEW程式的控制,整合為生物分子自動純化回收之電泳系統。 一開始的系統測試上,先以帶有負電性的DNA追蹤染劑為實驗樣本。因為DNA追蹤染劑在設計時即對應到不同長度的DNA片段,故可以此模擬DNA的實驗,進行純化回收的步驟。從實驗結果可看出,三條帶有不同顏色的染劑被各自回收到相異的緩衝溶液槽中。經由計算後得到回收率介於56%到88% 之間,證實此系統確實能執行自動純化回收的動作。因此,我們繼續以DNA樣本及PCR產物進行即時性生物感測實驗,結果顯示系統一樣能偵測到樣本訊號,並進行純化回收的動作,回收率介於40%到60% 之間,且無汙染的問題發生。 由以上結果可以得知,本研究開發的多端點膠體電泳裝置確實能進行生物分子的純化及回收動作,未來若能將其模組化或發展成可攜式即時檢測儀器,必定能為從事生物領域的研究人員節省許多時間及寶貴人力,獲得更高的實驗效率。 Biomolecule purification is an important step for biochemical experiments and there is an increasing need to develop automated purification system. For this reason, automated purification of biomolecules by multiterminal gel electrophoresis is investigated in this study. A PMMA-based, multiterminal electrophoresis device with dimensions of 50 mm x 100 mm x 10 mm was fabricated by machining in this work. The device featured a central channel for electrophoresis separation and two crossing channels for changing electrophoresis direction. The two LED/photodiode sensors were integrated with the device for automatic sample recovery. For demonstration, a mixture of three negatively charged dyes was tested in experiments of separation and purification. It was demonstrated that the dyes could be transported to different terminals sequentially within 30 min. Also, it was found that the dye recovery rate, ranged between 56% and 84%, was dependent on the electrophoresis distance: The longer path the dye traveled, the less the recovery rate was obtained. In addition to the dye molecules, automated separation of DNA fragments were done by a slight modification of the above device format. Accordingly, it is expected that the automated platform based on electrophoresis purification of biomolecules can be applied to on-chip screening and recovery applications. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37103 |
全文授權: | 有償授權 |
顯示於系所單位: | 生物機電工程學系 |
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