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研究生:何承錚
研究生(外文):Cheng-Cheng Ho
論文名稱:DNA探針引子間距長度對雜交與生物辨識的影響
論文名稱(外文):Effect of DNA Probe Spacer Length on Hybridization and Biorecognition
指導教授:胡文品
指導教授(外文):Wen-Pin Hu
口試委員:胡文品陳文逸陳玉菁
口試委員(外文):Wen-Pin HuWen-Yih ChenJui-Chi Chen 
口試日期:2012-07-23
學位類別:碩士
校院名稱:亞洲大學
系所名稱:生物與醫學資訊學系碩士班
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:52
中文關鍵詞:表面電漿共振核酸適體spacer間距長度凝血酶雜交DNA
外文關鍵詞:SPRaptamerspacer lengththrombinhybridizationDNA
相關次數:
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  • 下載下載:26
  • 收藏至我的研究室書目清單書目收藏:0
近年來生物科技發展趨勢越來越走向於基因治療、基因改質相關議題等,而這些相關的研究對於一個良好的生物感測器上也就顯得極為重要。在本研究中是使用表面電漿共振(Surface Plasmon Resonance, SPR)為一良好的生物感測器,先分析 Thiolated DNA 與MCH 在何種比例下的雜交反應量為最多,找出最佳 DNA 與 MCH 比例,再去探討具有不同間距(Spacer)長度的DNA做為探針使用時,對於量測效果的影響,去加以了解何種間距長度的DNA探針,可以提供較佳的雜交反應結果,產生最大的訊號量。此外,在本研究中也針對實驗室自組的表面電漿共振儀器進行測試,並使用此儀器進行凝血酶(Thrombin)與其核酸適體(Aptamer)結合反應的量測實驗。使用的兩種核酸適體(Aptamer)具有不同的間距長度,從實驗中再加以了解核酸適體之間距長度對於凝血酶與其結合反應的影響。
In the past few years, gene therapy, genetic modification and other related issues are the focuses in development of biotechnology. A good biosensor is also important to be used in some relevant biomedical research. In this study, we used the surface plasmon resonance (SPR) biosensors, a kind of biosensors, to perform experiments. Firstly, we studied how to prepare an optimal modification surface by using thiolated DNA and MCH. Then, we investigated the effect of using DNA probes with different spacer lengths on the DNA hybridization reactions. We use some DNA probes have 10, 20 or 26 continuous thymine bases at 5’ end. The hybridization efficiency of DNA on different surfaces of DNA chips was evaluated by a surface plasomon resonance (SPR) imaging. The experimental results indicate that the DNA probe with 10-thymine bases spacer shows the best hybridization efficiency. Besides, we used our home-made SPR instrument to study the interactions between the thrombin and two aptamers. One of these two aptamers had 10 continuous thymine bases at 5’ end; another one didn’t have any spacer at 5’ end. The result indicates that the aptamer had a spacer immobilized on sensor surface can interact with the thrombin better.
摘要 I
Abstract II
目錄 III
圖目錄 VII
第一章 緒論 1
1.1 前言 1
1.2 研究動機與目的 2
1.3 論文結構 3
第二章 文獻回顧 4
2.1生物感測器 4
2.2 表面電漿共振生物感測器 5
2.2.1 基本原理 6
2.2.1.1 Otto 組態 8
2.2.1.2 Kretschmann 組態 9
2.2.2 感測應用 10
2.3 DNA雜交反應 11
2.3.1 DNA雜交原理 12
2.3.2影響DNA雜交反應的因素 14
2.4 自我聚集單分子層的成膜方式 15
2.4.1 物理性成膜(Langmuir Blodgett薄膜) 15
2.4.2化學反應成膜 16
2.5 影響自組單層膜因素 19
2.5.1 反應溫度之影響 20
2.5.2 硫醇分子濃度之影響 20
2.5.3 分子碳鏈段長短之影響 20
2.6 抗蛋白質吸附的自我聚集單分子層 21
2.7 凝血酶與核酸適體之結合反應 21
第三章 材料與方法 24
3.1 化學藥品列表 24
3.2 儀器設備 26
3.2 DNA序列 27
3.2.1 用於探討DNA探針間距長度之序列 27
3.2.2 與凝血酶反應之核酸適體序列 27
3.3 緩衝溶液配置 28
3.4 SPR影像系統 29
3.5 微陣列晶片製作方式 30
3.5.1 DNA溶液配置 31
3.5.2 生物晶片製備 31
3.5.3 點片程序 31
3.5.4 SPR流體檢測 32
3.7 自組SPR量測系統 34
3.8 校正用溶液配置方式 36
3.9 凝血酶與具備兩種不同間距長度的核酸適體之實驗設計 37
第四章 結果與討論 39
4.1 使用不同比例的DNA與硫醇溶液製備DNA陣列於雜交反應的比較 39
4.2 提供最佳雜交反應的DNA探針間距長度 40
4.3 自組SPR儀器的校正數據 41
4.4凝血酶與具備兩種不同間距長度的核酸適體之結合反應比較 43
第五章 結論 46
5.1 研究的發現 46
5.2 未來展望 47
第六章 參考文獻 49


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