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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝之真(Chih-Chen Hsieh) | |
dc.contributor.author | Chin-An Lee | en |
dc.contributor.author | 李晉安 | zh_TW |
dc.date.accessioned | 2021-06-16T13:04:22Z | - |
dc.date.available | 2018-08-23 | |
dc.date.copyright | 2013-08-23 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-05 | |
dc.identifier.citation | 1. Turner, P.C., et al., 分子生物學速成. 臺北市: 合記書局總經銷, 2002.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61500 | - |
dc.description.abstract | 本研究的目的為開發一拉伸DNA平台,並將其應用於基因圖譜技術。現行的拉伸DNA技術大多利用侷限空間迫使DNA朝無侷限空間展開而達到拉伸效果,其中以奈米通道拉伸法最受大家關注,其優點在於DNA的伸長率佳且DNA的伸長型態為其平衡狀態,利於長時間下的觀察及分析,因此較適合用於基因圖譜的檢測。然而,奈米通道在製作上及操作上皆具有困難度,以整體成本及效益來說,不利於醫療單位的使用。因此我們希望發展一穩定拉伸DNA的平台,且同時具有極佳的伸長效果且低製作成本。
我們沿用侷限空間的概念,將帶負電的DNA吸附於帶正電的脂雙層上,使DNA呈現二維運動行為,並以圖案脂雙層使DNA吸附後於一維方向自發伸展,其中伸展機制是藉由降低系統位能來克服DNA型態變化所產生的熵損失,使DNA伸展型態為系統中最低自由能狀態,即其平衡狀態。然而,實驗上發現氯仿存在於脂雙層內才可觀察到DNA伸展現象,且氯仿含量與DNA伸長率呈正相關,但目前仍無法完全了解其作用。實驗結果顯示lambda DNA的伸長率可達到73%,其相當接近奈米通道拉伸法的極限,本實驗平台之成本遠低奈米通道,較利於基因圖譜檢測應用,有助於未來實際運用於醫療單位。此外,我們也發現不同大小的DNA於相同實驗條件下的伸長率非常接近,此特性能夠將DNA長度轉為基因圖譜檢測特徵之一,使此法應用於基因圖譜檢測能更加準確。 | zh_TW |
dc.description.abstract | The purpose of this study is to develop a new platform for stretching DNA, and then to apply it to the application of DNA gene mapping. Among the recently developed technologies for DNA gene mapping, using nanochannel to confine and therefore to stretch DNA is the most promising one. DNA confined in nanochannel spontaneously extends to nearly its contour length, and the positions of specific sequences can be readily determined. However, the nanochannel is not only expensive to fabricate but also difficult to operate. Thus, a more efficient, lower-cost, and easy-to-use DNA-stretching platform is much anticipated.
Following the concept of confinement, we adsorb DNA on a patterned two-dimensional plane built by cationic lipid bilayers. The pattern consists of periodic shallow trenches, and DNA spontaneously gathers and unravels along the root of the trench wall. The phenomenon is controlled by the competition between the electrostatic interaction and conformational entropy of DNA. However, we also discovered that the existence of chloroform in our system strongly affects the DNA behavior. Although the mechanism is yet to be investigated, the degree of DNA extension was found to increase with chloroform content. For both T4 and Lambda DNA, their extensions were found very close and can be more than 70% of their contour lengths, comparable to the value obtained in nanochannels. Therefore, the platform developed in this study can be considered as a cheaper alternative of nanochannel. The current platform will be tested for gene mapping in the very near future. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T13:04:22Z (GMT). No. of bitstreams: 1 ntu-102-R00524078-1.pdf: 6143806 bytes, checksum: 222b64ee7c9f0ced8b89bc4a740fc13a (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 目錄
摘要 ………………………………………………………………………………I Abstract ……………………………………………………………………………...II 目錄 …………………………………………………………………………….III 圖目錄 ……………………………………………………………………………...V 表目錄 …………………………………………………………………………….XI 符號表 ……………………………………………………………………………XII 希臘符號表 XII 第1章 緒論 1 1.1 前言 1 1.2 基因圖譜 1 1.2.1 DNA高分子之介紹 1 1.2.2 基因圖譜技術之發展 5 1.3 研究動機與目的 9 第2章 文獻回顧 10 2.1 DNA高分子之物理性質 10 2.2 DNA拉伸技術 11 2.2.1 栓扯拉伸法 11 2.2.2 分子梳拉伸法 13 2.2.3 流電場梯度拉伸法 15 2.2.4 侷限拉伸法 18 2.3 DNA吸附於脂雙層上之行為 21 2.3.1 脂質(Lipids) 21 2.3.2 脂雙層(Supported Lipid Bilayers,SLBs) 24 2.3.3 螢光漂白後回復技術(Fluorescence Recovery After Photobleaching,FRAP) 29 2.3.4 DNA於脂雙層上之行為 33 2.3.5 DNA於圖案脂雙層上自發展開之行為 37 第3章 實驗設計 43 第4章 實驗設備與步驟 46 4.1 儀器設備 46 4.2 實驗藥品 48 4.3 實驗方法 50 4.3.1 圖案玻片之製程 50 4.3.2 架設脂雙層 53 4.3.3 DNA溶液之製備 56 4.3.4 觀察DNA於脂雙層上 57 第5章 實驗結果與討論 59 5.1 圖案玻片之製程結果 59 5.2 DNA於脂雙層上自發展開之結果 63 5.2.1 熱力學初步分析 63 5.2.2 DNA吸附於脂雙層後自發展開之過程 68 5.2.3 DNA沿側壁內角伸展之現象 71 5.2.4 DNA自發伸展機制 75 5.2.5 DNA伸長量之結果及討論 85 5.2.6 其他結果 92 5.3 結果比較及討論 95 第6章 結論 100 第7章 參考文獻 102 | |
dc.language.iso | zh-TW | |
dc.title | DNA於脂雙層上自發展開之研究 | zh_TW |
dc.title | Research on the Spontaneous Unraveling of DNA on Lipid Bilayer | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 趙 玲(Ling Chao),童世煌(Shih-Huang Tung),林宏殷(Hung-Yin Lin) | |
dc.subject.keyword | 去氧核醣核苷,酸,脂雙層, | zh_TW |
dc.subject.keyword | DNA,Lipid Bilayer, | en |
dc.relation.page | 105 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-08-05 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
顯示於系所單位: | 化學工程學系 |
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