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研究生:梁柏榮
研究生(外文):Bo-Rong Liang
論文名稱:電子式基因序列偵測晶片之原型
論文名稱(外文):A prototype of gene-chip based on electrical detection
指導教授:蔡章仁蔡章仁引用關係辛裕明
指導教授(外文):Jang-Zern TsaiYue-ming Hsin
學位類別:碩士
校院名稱:國立中央大學
系所名稱:電機工程研究所
學門:工程學門
學類:電資工程學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:93
中文關鍵詞:改質阻抗導納奈米粒子生物晶片
外文關鍵詞:impedanceDNAmodificationbiochipnanoparticle
相關次數:
  • 被引用被引用:2
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本研究的目的是研討電性偵測用於微米等級的基因晶片之檢測。使用半導體製程技術,在晶片上製作梳狀微電極,此微電極類似超大型積體電路的電容器構造,兩個電極為一組,每一組電極中間的間距為數微米,使用表面化學改質技術改質電極中間的區域,先將單股的寡核苷酸序列固定上去,之後進行雜交反應。雜交反應完成後,再將奈米金粒子與寡核苷酸分子的最頂端的端點的硫鍵結,於是奈米金粒子可當成雜交反應的標記。利用此程序可避免複雜的化學處理步驟,而且不需要奈米級製程,只需用到微米級製程就可以做到夠小的電極間距,達到有效偵測。
傳統的 DNA 雜交偵測方法有一些缺點,如螢光標記的信號會隨時間衰減,放射線標記傷害人體,而且偵測這些信號必須使用昂貴的偵測裝置。本研究使用直流電與交流電量測微電極,發現雜交反應的發生 ( 奈米金粒子鍵結 ) 會使量測到的信號與沒有雜交反應的信號有差異,進而利用這些信號達到檢測的目標,便可避開傳統的偵測法的缺點。實驗成功後研製一組簡單的量測電路,在量測時不必使用昂貴的儀器,可降低成本,還可以大量偵測且偵測自動化。
雜交反應的偵測是基因晶片的重點技術,本研究的偵測技術可應用在基因晶片的基因表達、篩檢、及比對等研究,也可以應用在病原體基因檢測、基因表現比較、基因突變分析、基因序列分析等領域。
In this research, we make comb-shape electrodes using semiconductor process. One pair of electrodes is two electrodes, and the distance between them is several micro-meter. Electrodes are like capacitance structure of VLSI. The surface was treated by surface chemical modification and was immobilized with single strand oligonuceotide, then carry out hybridization. After the hybridization, the top-end of DNA was bond with gold nanoparticle. The silver-enhancer and gold muiltilayer deposition can be avoided if the process was used. Furthermore, the good detection was discovered and only use of UV exposure without E-beam lithography.
There are some disadvantages of tradition detection methods, for example, fluorescence signal decay with time, and human body was hurt by radioactive labeling. The research avoid disadvantages of traditional detection methods, the detection was implemented by DC measurement and AC measurement on micro-electrodes. After the binding of gold nanoparticle, the change of admittance value was discovered. The detection of hybridization can be represented by the admittance value.
Hybridization detection is the key technique of gene chip, the detection method of the research can be applied on gene expression, and quantification of expressed genes, differentistion of expression genes, mutations, identification of sequence, evaluation of specific DNA-binding proteins or molecules.
論文摘要------------------------------------------------------------------I
Abstract-------------------------------------------------------------------II
目錄----------------------------------------------------------------------III
圖目錄--------------------------------------------------------------------V
表目錄-------------------------------------------------------------------IX

第一章 緒論--------------------------------------------------------1
1.1 前言----------------------------------------------------------------1
1.2 動機和目標-------------------------------------------------------4
1.3 文獻回顧----------------------------------------------------------5
1.3.1 國內外有關本研究之研究情況-------------------------5
1.3.2 參考文獻探討---------------------------------------------6
1.4 論文架構----------------------------------------------------------8

第二章 研究方法與原理----------------------------------------10
2.1 DNA 分子簡介-------------------------------------------------10
2.2 基因晶片的檢測原理------------------------------------------17
2.3 基因晶片的檢測流程------------------------------------------27

第三章 實驗方法與步驟----------------------------------------29
3.1 基因晶片之微電極設計與製程---------------------------------------29

3.1.1 微電極圖案設計與製程規劃--------------------------------29
3.1.2 實驗儀器設備--------------------------------------------------32
3.1.3 微電極製程步驟-----------------------------------------------36
3.2 DNA之固定化與雜交反應實驗---------------------------------------40
3.2.1 實驗藥品與寡核苷酸序列----------------------------------------40
3.2.2 儀器儀器設備----------------------------------------------------49
3.2.3 實驗目的---------------------------------------------------------50
3.2.4 實驗方法---------------------------------------------------------53
3.3 電性量測與SEM觀測---------------------------------------------------60
3.3.1 量測之儀器設備------------------------------------------------60
3.3.2 電性量測實驗---------------------------------------------------63
3.3.3 SEM觀測實驗--------------------------------------------------64
3.4 量測電路與基因晶片之整合-------------------------------------------64
3.5 非相等長度電極的實驗-------------------------------------------------68

第四章 結果與討論------------------------------------------------------72
4.1 電性量測結果與討論-------------------------------------------------72
4.2 奈米金粒子之分佈情形-------------------------------------------------76
4.3 量測電路與基因晶片之整合的結果與討論-------------------------82
4.4 非相等長度電極的實驗之結果與討論--------------------------------83

第五章 結論與未來展望-------------------------------------------85

參考文獻------------------------------------------------------------88
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