請用此 Handle URI 來引用此文件：
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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 范士岡 | |
| dc.contributor.author | An-Te Chen | en |
| dc.contributor.author | 陳安得 | zh_TW |
| dc.date.accessioned | 2021-06-16T09:57:54Z | - |
| dc.date.available | 2022-02-08 | |
| dc.date.copyright | 2017-02-08 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-12-14 | |
| dc.identifier.citation | [1] E. T. Lagally, C. A. Emrich and R. A. Mathies, “Fully integrated PCR-capillary electrophoresis microsystem for DNA analysis,” Lab on a Chip, vol. 1, pp. 102-107, 2001.
[2] R. M. Guijt, A. Dodge, G. W. K. van Dedem, N. F. de Rooija and E. Verpoort , “Chemical and physical processes for integrated temperature control in microfluidic devices,” Lab on a Chip, vol. 3 pp. 1-4, 2003. [3] V. Srinivasan, V. K. Pamula and R. B. Fair, “An integrated digital microfluidic lab-on-a-chip for clinical diagnostics on human physiological fluids,” Lab on a Chip, vol. 4, pp.310-315, 2004. [4] C-J. Kim, “Microfluidics using the surface tension force in microscale,” SPIE conf. on micromaching and microfabrication, Santa Clara, 2000, pp. 44-55. [5] P. Neuzil, J. Pipper and T. M. Hsieh, “Disposable real-time microPCR device： lab-on-a-chip at a low cost,” Lab on a Chip, vol. 2, pp. 292-298, 2006. [6] M. A. Northru, C. Gonzalez, D. Hadley, R.F. Hills, P. Landre, S. Lehew, R. Saikil, J. J. Sninskyll, R. Watson and R. Watson, Jr., “A MEMS-based Miniature DNA analysis system,” The 8th International Conf. on Solid-State Sensors and Actuators, Sweden, 1995, pp. 764-767. [7] J. Berthier, Microdrops and Digital Microfluidics, 1st ed. pp. France: William Andrew Inc., 2008, pp. 323-326. [8] D. J. Boles, J. L. Benton, G. J. Siew, M. H. Levy, P. K. Thwa, M. A. Sandahl, J. L. Rouse, L. C. Perkins, A. P. Sudarsan, R. Jalili, V. K. Pamula, V. Srinivasan, R, B, Fair, P. B. Griffin, A. E. Eckhardt and M. G. Pollack, “Droplet-based pyrosequencing using digital microfluidics” Analytical Chemistry, vol. 83, pp. 8349-8447, 2011 [9] T. Chen, Y. Jia, C. Dong, J. Gao, P. Mak and R. P. Martins, “Sub-7-second genotyping of single-nucleotide polymorphism by high-resolution melting curve analysis on a thermal digital microfluidic device,” Lab on a Chip, vol. 16, pp. 743-752, 2016. [10] K. A. Connors, Chemical Kinetics： The Study of Reaction Rates in Solution, John Wiley & Sons, 1990, pp. 3-8. [11] Y. C. Chung, M. S. Jan, Y. C. Lin, J. H. Lin, W. C. Cheng and C. Y. Fan, “Microfluidic chip for high efficiency DNA extraction,” Lab on a Chip, vol. 4, pp. 141-147, 2004 [12] W. C. Hui, L. Yobas, V. D. Samper, C. K Heng, S. Liwa, H. Ji, Y. Chen, L. Cong, J. Li and T. M. Limb, “Microfluidic systems for extracting nucleic acids for DNA and RNA analysis,” Sensors and Actuators, vol. 133, pp. 335-339, 2007. [13] M. Mahalanabis, H. Al-Muayad, M. D. Kulinski, D. Altmanb and C. M. Klapperichac, “Cell lysis and DNA extraction of gram-positive and gram-negative bacteria from whole blood in a disposable microfluidic chip,” Lab on a Chip, vol. 9, pp. 2811-2817, 2009 [14] M. Karle, J. Miwa, G. Czilwik, V. Auw?arter, G. Roth, R. Zengerleabd and F. von Stettenab, “Continuous microfluidic DNA extraction using phase-transfer magnetophoresis, ” Lab on a Chip, vol. 10, pp. 3284-3290, 2010 [15] R. Sista, Z. Hua, P. Thwar, A. Sudarsan, V. Srinivasan, A. Eckhardt, M. Pollack and V. Pamula, “Development of a digital microfluidic platform for point of care testing,” Lab on a Chip, vol. 8, pp. 2091-2104, 2008. [16] 洪秉毅， | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60128 | - |
| dc.description.abstract | 本文是利用數位微流體 (Digital microfluidic，DMF) 技術整合DNA萃取和即時聚合酶鏈鎖反應 (Real-time polymerase chain reaction，qPCR) 於晶片內。為了使point-of-care更加普及化，未來必須發展出一個個人裝置在不需要臨床人員下，能夠在任何時候和任何地方都可以即時檢測疾病，隨時都能掌握自己的健康狀況，最關鍵的技術就在DNA萃取和qPCR。在此晶片上可利用交流電產生介電濕潤 (Electrowetting on dielectric，EWOD) 來移動人體血液跟生物試劑套件混合來萃取血液裡的DNA，在100 nL的血液裡萃取出4.45 ng/μL濃度的DNA。跟傳統的DNA萃取比較，萃取時間減少75 %的反應時間，從120分鐘縮短至30分鐘，試劑消耗減少99.95 %的體積，從4222 | zh_TW |
| dc.description.abstract | This thesis reports the implementation of DNA extraction and real-time PCR (qPCR) on a digital microfluidic (DMF) device. We aim to develop a personalized point-of-care device for molecular diagnosis from human whole blood and commercial reagents kits driven by electrowetting-on-dielectric (EWOD) on a DMF device for DNA extraction and qPCR. The results from on-chip DNA extraction protocols were validated and quantified. In comparison wiht the traditional DNA extraction procedures in tubes, our on-chip extraction starting from a 100 nL whole blood obtained 52.8 % DNA concenttion at 4.45 ng/µL, required 25 % reaction time (from 120 min to 30 min), consumed 0.05 % volume of the entire reagents (from 4222 | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T09:57:54Z (GMT). No. of bitstreams: 1 ntu-105-R03522114-1.pdf: 4148207 bytes, checksum: 23c6eeb5efae4875c79a154cbdac318c (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 口試委員會審定書 #
中文摘要 i ABSTRACT ii 誌謝 iii 目錄 vi 圖目錄 vii 表目錄 xi 第一章 緒論 1 1.1 前言 1 1.2 文獻回顧 3 1.2.1 DNA萃取微流體晶片 3 1.2.1.1 DNA萃取連續微流體晶片 4 1.2.1.2 DNA萃取數位微流體晶片 6 1.2.2 qPCR及PCR微流體晶片 8 1.2.2.1 qPCR及PCR連續微流體晶片 8 1.2.2.2 qPCR及PCR數位微流體晶片 11 1.3 研究目標與動機 16 第二章 理論與概念設計 17 2.1 操控-介電濕潤之理論 17 2.2 加熱-數位微流體晶片加熱之理論 23 2.2.1介質式加熱 23 2.2.2電阻式加熱 25 2.3 DNA萃取 26 2.3.1 DNA萃取方法介紹 26 2.3.2磁珠式DNA萃取 28 2.4 PCR原理 31 2.5 qPCR原理 35 第三章 元件製程與實驗架設 36 3.1 晶片設計與製作 36 3.1.1 光罩設計概念 36 3.1.2 晶片製作流程 40 3.2 實驗架設與儀器介紹 44 3.2.1 儀器功能介紹 44 3.2.2 整體實驗的架設 47 3.3 實驗材料介紹 48 3.3.1 Agencourt Genfind v2 Kits 48 3.3.2 PCR和qPCR試劑 50 第四章 實驗和結果分析 51 4.1 數位微流體DNA萃取晶片的研發 51 4.1.1血和DNA萃取試劑的接觸角測量 51 4.1.2數位微流體晶片上的液體操作 53 4.1.3數位微流體晶片上的DNA萃取–磁珠聚集問題 56 4.1.4優化後數位微流體晶片上的DNA萃取 64 4.1.5晶片上DNA萃取產物定量分析 72 4.2 數位微流體qPCR晶片的研發 75 4.2.1 傳統PCR 76 4.2.2 初步qPCR溶液加熱測試 77 4.2.3 量測溫度校正 79 4.2.4 qPCR溶液驅動分析 82 4.2.5溫度分布和溫控穩定性之分析 84 4.2.6晶片上qPCR 88 第五章 結論和未來展望 94 5.1 結論 94 5.2 未來展望 95 參考文獻 96 | |
| dc.language.iso | zh-TW | |
| dc.subject | 生物實驗室晶片 | zh_TW |
| dc.subject | DNA萃取 | zh_TW |
| dc.subject | 即時聚合?鏈鎖反應 | zh_TW |
| dc.subject | 數位微流體 | zh_TW |
| dc.subject | Electrowetting-on-dielectric | en |
| dc.subject | Lab-on-chip | en |
| dc.subject | Digital microfluidics | en |
| dc.subject | DNA extraction | en |
| dc.subject | qPCR | en |
| dc.title | DNA萃取和即時聚合酶鏈鎖反應於數位微流體晶片 | zh_TW |
| dc.title | DNA Extraction and Real-time Polymerase Chain Reaction on Digital Microfluidic Chip | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 盧彥文,謝之真,楊燿州,張鈺 | |
| dc.subject.keyword | 生物實驗室晶片,DNA萃取,即時聚合?鏈鎖反應,數位微流體, | zh_TW |
| dc.subject.keyword | Lab-on-chip,Electrowetting-on-dielectric,Digital microfluidics,DNA extraction,qPCR, | en |
| dc.relation.page | 101 | |
| dc.identifier.doi | 10.6342/NTU201603805 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-12-15 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 機械工程學研究所 | zh_TW |
| 顯示於系所單位： | 機械工程學系 | |
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|---|---|---|---|
| ntu-105-1.pdf 未授權公開取用 | 4.05 MB | Adobe PDF |
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