博碩士論文 103324003 詳細資訊




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姓名 曾柏偉(Po-Wei Tseng)  查詢紙本館藏   畢業系所 化學工程與材料工程學系
論文名稱 矽奈米線場效電晶體多點之核酸檢測研究
(Development of multi-point immobilization system on silicon nanowire field effect transistor)
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摘要(中) 近年來有各種生物標記物( biomarkers )的檢測方法被發展與推廣。這些檢測方法包括利用光學、電學、質量等運作原理的生物感測器( biosensors )來偵測DNA、RNA、蛋白質等所謂之生物標記分子。在眾多的生物感測器中,因為矽奈米線場效電晶體相較於其他類別的生物感測器有著超靈敏、具選擇性、即時且免螢光標定的優勢,因此被廣泛的研究與發展,也使得矽奈米線場效電晶體在未來醫學診斷方面有很大的應用潛力。現今矽奈米線場效電晶體的生物分子固定化流程大多採用整片式固定法( all area modification ),然而以這種方式固定上生物受體只能檢測一種生物標記物。
為了解決這個問題,我們開發了「多點固定化系統」。此系統可以將多種生物受體( bioreceptors )溶液點在晶片上的每個元件上,若矽奈米線場效電晶體有更多種生物探針固定於其上,我們就能在同一個晶片上檢測多種生物標記物就是所謂的高通量檢測。相較於整片式固定法,節省成本與實驗的時間是一個很大的優勢。
在本研究,我們首先探討DNA在不同鹽離子濃度下對雜交的影響,接下來我們再探索探針溶液中的甘油比例與探針濃度於多點固定化系統的實驗最佳條件,最後我們嘗試在同一片晶片上固定不同種類的生物探針來進行核酸檢測。
摘要(英) In recent years, various biomarker detection methods have been invented and studied. Optical, mass, electrical and etc… biosensors are common for monitoring DNA, RNA, protein, and so on. Among them, Silicon nanowire field effect transistor (SiNW-FET) is the one that has been extensively studied and developed due to its distinguished advantages over the other biosensors. SiNW-FET is ultrasensitive, selective, real-time and label-free detection platform. Because of these features, SiNW-FET has a great potential in the future of clinical diagnostics. In the current process of SiNW-FET immobilization, we mostly use all area modification. However, if we use this method to immobilize bioreceptors on SiNW-FET, the device can detect only one kind of biomarker. To solve this problem, we develop a technique called “multi-point immobilization system”. Multi-point immobilization system can overcome the above-described problems because this system can spot many kinds of bioreceptor solution on each device. If there are more bioreceptor probes on SiNW-FET, we can detect more categories of biomarker targets on same device. Saving costs and experimental time are comparative advantage with all area modification. In this research, first we investigate the effects of salt ion concentration on hybridizing. Then we explore the best ratio of glycerol in probe solution and optimal probe concentration in multi-point immobilization system. Finally we try to immobilize different probe on same device to measure electrical signal.
關鍵字(中) ★ 場效電晶體
★ 核酸分子
★ 奈米線
★ 微陣列
關鍵字(英) ★ field effect transistor
★ nucleic acids
★ nanowire
★ microarray
論文目次 摘要 i
Abstract iii
誌謝 iv
目錄 v
圖目錄 viii
表目錄 xiv
第一章 緒論 - 1 -
第二章 文獻回顧 - 3 -
2.1 場效電晶體生物感測器 - 3 -
2.2 德拜長度對電性檢測的限制 - 10 -
2.3 核酸適體簡介 - 13 -
2.4 核酸類似物 - 18 -
2.4.1 肽核酸 - 18 -
2.4.2 鎖核酸 - 19 -
2.4.3 磷酸根甲基化去氧核醣核酸 - 20 -
2.5 晶片表面改質與固定化 - 22 -
2.6 生物晶片與微陣列技術 - 26 -
2.7 檢測訊號之放大技術 - 32 -
第三章 實驗藥品、儀器設備與方法 - 34 -
3.1 實驗藥品 - 34 -
3.1.1 化學品 - 34 -
3.1.2 實驗溶液 - 35 -
3.1.3 實驗核酸序列 - 36 -
3.2 儀器設備 - 36 -
3.3 實驗方法 - 37 -
3.3.1 R-18 RNA aptamer合成 - 37 -
3.3.2 PCR產物之cleaning up - 38 -
3.3.3 轉錄R-18 RNA aptamer - 39 -
3.3.4 R-18 RNA aptamer電泳測試 - 40 -
3.3.5 FET晶片固定化 - 40 -
3.3.6 FET電性測量 - 42 -
第四章 結果與討論 - 43 -
4.1 DNA與nDNA之雜交鑑定 - 43 -
4.2 DNA 於不同鹽離子濃度的雜交測試 - 45 -
4.3 探針溶液之甘油含量對雜交效率的影響 - 49 -
4.4 探針溶液濃度對雜交效率的影響 - 53 -
4.5 比較nDNA與DNA探針之雜交效率 - 56 -
4.6 固定多點核酸探針之雜交檢測 - 60 -
4.7 R18 RNA適體之訊號放大實驗 - 65 -
4.8 實驗晶片良率統計 - 77 -
第五章 結論 - 78 -
第六章 參考文獻 - 80 -
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指導教授 陳文逸(Wen-Yih Chen) 審核日期 2016-7-22
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