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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82137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 | Shao-Yang Huang | en |
| dc.contributor.author | 黃紹洋 | zh_TW |
| dc.date.accessioned | 2022-11-25T05:36:40Z | - |
| dc.date.available | 2023-10-06 | |
| dc.date.copyright | 2021-11-02 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-10-07 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82137 | - |
| dc.description.abstract | 在分子分形時，難免會有疏漏而造成線形與環形DNA混合，也因此產生分離不同構型DNA的需求。在現行應用中，最常見且傳統之分離方法為凝膠電泳，然耗時長為其缺點；而最近利用微流道分離不同構型DNA的研究中，則多以絕緣式介電泳來分離，然而各種不同設計皆有其侷限。在本研究中，我們則嘗試在微流道中利用正向應力或介電泳力分離線型及環形之DNA。首先，我們已經在先前的研究中實現以正向應力(normal stress)來分離不同長度之線型DNA，也因此欲探究以此裝置分離不同構型DNA之可能性。在利用介電泳力方面，則希望設計出有別以往絕緣式介電泳分離之新穎裝置。我們以λ-DNA(48.5 kbp)和Charomid 9-42 DNA(42.2 kbp)分別代表線形與環形做為分離之對象。 在以正向應力分離之實驗部分，因線形與環形DNA之鬆弛時間差距不大，造成兩者正向應力差別有限，導致分離效果差於預期。為改善分離效率，我們使用鞘流將DNA於裝置入口處先集中，使正向應力的效應放大，確實能夠使線形DNA傾向往側邊出口流動，而環形DNA則傾向沿著裝置中央流動，達到較佳的分離。 我們亦嘗試以交流電引發DNA負介電泳，以搭配正向應力來更進一步提升分離DNA的效率。儘管從文獻中測量DNA與溶液介電係數之結果可以推測DNA非常難產生負介電泳力之現象，但仍有許多關於DNA負介電泳之參考文獻。也因此我們嘗試文獻上不同配方之緩衝溶液，並改變頻率範圍，然皆未發現明顯負介電泳之現象。此外，實驗過程中不論於PDMS微流道或是玻璃微流道內，皆觀測到明顯之電流熱效應，此效應造成之DNA運動與負介電泳非常近似，然電流熱效應並未被上述任何一篇文獻所考慮，所以我們認為文獻中電關於DNA負介電泳現象很可能是由電流熱效應所產生。 因並未發現負介電泳之現象，我們轉為以正介電泳力分離DNA，並設計出有別於以往以介電泳力分離之裝置。在新穎以絕緣式介電泳分離之實驗中，除了λ-DNA和Charomid 9-42 DNA，我們亦加入T4 DNA (165.6 kbp)，以同時探討於此裝置分離不同長度DNA之可能性。在此裝置中，我們確實觀測到不同DNA在相同介電泳條件下偏移程度有差異，但此設計仍需進一步優化使其分離效果提升。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T05:36:40Z (GMT). No. of bitstreams: 1 U0001-0610202102024400.pdf: 9350790 bytes, checksum: 7f45626d7849e4881094e81bbc784f4b (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 論文口試委員審定書 I 致謝 III 摘要 IV Abstract VI 目錄 VIII 圖目錄 XI 表目錄 XXXIII 第 1 章 、緒論 1 1.1 前言 1 1.2 研究動機 1 第 2 章 、文獻回顧 3 2.1 DNA簡介 3 2.1.1 DNA的化學結構 4 2.1.2 DNA的高分子性質 7 2.1.3 DNA與染劑分子 8 2.1.4 DNA的構型 10 2.1.5 線與環DNA的物理性質 11 2-2 DNA常見的電動力學 14 2.2.1 電雙層 15 2.2.2 電泳 15 2.2.3 電滲流 16 2.2.4 電流熱效應 17 2.3 電流體動力不穩定性(electrohydrodynamic instability) 18 2.4 分離原則與分析方法 21 2.4.1 分離解析度 22 2.4.2 分離效率 24 2.4.3 位移效率 25 2.5 現行DNA分離技術 28 2.5.1 凝膠電泳(gel electrophoresis，GEP) 28 2.5.2 毛細管電泳(capillary electrophoresis，CE) 29 2.5.3 微流道電泳(microchannel electrophoresis) 30 2.5.4 線與環之DNA於之分離 38 2.6 介電泳 40 2.6.1 介電泳力和電容率及電導率之關係 42 2.6.2 介電泳於微流道分離之應用 47 2.6.3 介電泳於DNA之應用 59 2.7 新穎正向應力分離DNA之裝置 73 2.7.2 結合正向應力之側向分離通道 76 2.7.3 影響正向應力之因素 78 2.8 研究目的與實驗設計構想 80 2.8.1.以正向應力分離環與線的DNA 80 2.8.2 結合負介電泳力與正向應力以側向分離DNA 85 2.8.3 開發新形絕緣式介電泳之裝置 88 第 3 章 、設備、材料與方法 92 3.1 實驗設備 92 3.2 實驗材料 95 3.3 通道製作 97 3.4 溶液配置 109 3.5 電場施加裝置 112 3.6 結果收集與分析 118 第 4 章 、結果與討論 121 4.1 以正向應力分離線與環之DNA 121 4.1.1 分叉出口對分離之影響 123 4.1.2 以集中流場分離之結果 131 4.2 負介電泳力之測試 144 4.2.1 不同濃度之PBS緩衝液 146 4.2.2 pH 8.0 NaOH 150 4.3 開發新穎之絕緣式介電泳分離裝置 160 第 5 章 、結論與未來展望 174 5.1 結論 174 5.2 未來展望 175 第 6 章 、參考文獻 176 | |
| dc.language.iso | zh-TW | |
| dc.subject | 環狀DNA分離 | zh_TW |
| dc.subject | 絕緣式介電泳 | zh_TW |
| dc.subject | 微流道 | zh_TW |
| dc.subject | 介電泳 | zh_TW |
| dc.subject | 正向應力 | zh_TW |
| dc.subject | insulated dielectrophoresis | en |
| dc.subject | Microchannel | en |
| dc.subject | circular DNA | en |
| dc.subject | DNA separation | en |
| dc.subject | normal stress | en |
| dc.title | 以正向應力及介電泳力在微流道中分離線形與環形DNA之研究 | zh_TW |
| dc.title | Separation of Linear and Ring DNA by Normal Stress and Dielectrophoresis Force in Microchannels | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 趙玲(Hsin-Tsai Liu),莊怡哲(Chih-Yang Tseng),周家復 | |
| dc.subject.keyword | 微流道,環狀DNA分離,正向應力,介電泳,絕緣式介電泳, | zh_TW |
| dc.subject.keyword | Microchannel,circular DNA,DNA separation,normal stress,insulated dielectrophoresis, | en |
| dc.relation.page | 180 | |
| dc.identifier.doi | 10.6342/NTU202103570 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-10-08 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
| dc.date.embargo-lift | 2023-10-06 | - |
| 顯示於系所單位： | 化學工程學系 | |
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