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研究生:邱馨慧
研究生(外文):Shin-Huei Chiou
論文名稱:奈米粒子對毛細管電泳DNA分離之影響及光合成法製備奈米金球
論文名稱(外文):The impacts of gold nanoparticles on DNA separation by capillary electrophoresis and photosynthesis of gold nanoparticles in the presence of proteins
指導教授:張煥宗張煥宗引用關係
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:化學研究所
學門:自然科學學門
學類:化學學類
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:100
中文關鍵詞:金奈米粒子毛細管電泳DNA
外文關鍵詞:capillary electrophoresisgold nanoparticlesDNA
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本論文主要分成兩部分,一是探討金奈米粒子對毛細管電泳DNA分離的影響;二則以光合成法在蛋白質存在下製備奈米金球。利用聚環氧乙烯﹙poly(ethylene oxide);PEO﹚以及金奈米粒子﹙gold nanoparticles;GNPs﹚在毛細管電泳系統中分離小片段DNA具有良好的再現性、高解析度以及快速分離之優點。本研究的第一部分進一步探討緩衝溶液之pH值、GNPs之濃度、染料分子與GNPs之間濃度的比例關係及鹽類的含量等因素對分離結果的影響。在pH 7.0-10.0範圍,以0.2% PEO﹙8M Da﹚及0.3X 56-nm GNPs﹙9.87 × 1012 particles/L﹚進行DNA markers V和VI混合物之電泳分離,發現GNPs在pH 9.0下對DNA的分離有較明顯的助益。固定pH 為9.0時,提高PEO與EtBr濃度則有助於小片段DNA的解析。雖然GNPs濃度增加時,會減少DNA與毛細管壁作用而使分離結果再現性佳,但是在固定PEO濃度與GNP/EtBr濃度比例之條件下,過多的GNPs存在反而使小於504 bp之DNA片段解析度變差。此外,研究中更進一步發現當在上述最佳條件中添加0.01 mM檸檬酸鹽﹙citrate﹚可使分離解析度增加及縮短分離時間。該條件並成母野峏顜硈t(7分鐘內)分離DNA Marker V(pBR322/ Hae III digest)、ψX 174 RF DNA-Hae III digest及lambda DNA Hind III digest之混合物(0.008-23.1 kbp)。第二部份則嘗試以光照法在蛋白質存在下合成GNPs。金奈米粒子的大小與反應速率受控於蛋白質與金離子的相對及絕對濃度,同時也受溶液pH值的影響。本研究中大小為7-12 nm的GNPs是在NaAuCl4及BSA的最終濃度為1 mM及10 μM時所得。依此比例,變換溶液pH值及環境溫度觀察奈米粒子變化情形,發現在pH 7.0條件下,GNPs的粒徑分佈較小且均勻。根據室溫下不照光時GNPs無法合成的結果,推論光為促使金離子還原的主要原因,而蛋白質則是防止GNPs聚集的保護劑。相較於傳統以檸檬酸鹽在加熱下還原金離子,此法可直接在PDMS上進行反應,且合成的GNPs以物理吸附方式吸附於PDMS表面,未來可應用於生物感測器的研究。
摘要 Ⅰ
目錄 Ⅲ
表目錄 Ⅶ
圖目錄 Ⅷ



第一章 毛細管電泳及奈米粒子應用概論 1
1.1 毛細管電泳歷史沿革 1
1.2 毛細管電泳理論基礎 5
1.2.1 電泳 5
1.2.2 電滲流與Zeta電位 6
1.2.3 分離解析度 8
1.2.4 分離效率 8
1.2.5 影響分離效率的因素 9
1.2.5.1 焦耳熱 9
1.2.5.2 進樣長度 10
1.2.5.3 溶質與管壁相互作用 11
1.3 毛細管電泳之分離模式 11
1.3.1 毛細管凝膠電泳(capillary gel electrophoresis,CGE) 12
1.3.2 聚合物溶液(polymer solution) 13
1.3.3 DNA的電泳分離機制 14
1.3.3.1 Ogston模型 15
1.3.3.2 Reptation模型 17
1.4 毛細管電泳之偵測系統 18
1.4.1 UV吸收法 18
1.4.2雷射激發誘導螢光 19
1.4.3質譜儀 20
1.5 毛細管電泳之DNA樣品分析之應用 21
1.5.1 DNA序列分析 21
1.5.2 點突變偵測 23
1.6 奈米科技簡介 24
1.6.1 奈米粒子的製備 27
1.6.2 奈米技術在分子生物與醫學領域的應用 29
1.7 研究動機 31
1.8 參考文獻 34
1.9 本章圖表 43
第二章 金奈米聚合物溶液對毛細管電泳之DNA分離影響因素探討 51
2.1 導論 51
2.2 實驗 54
2.2.1 毛細管電泳系統 54
2.2.2 雷射誘導螢光偵測系統 54
2.2.3 實驗試藥 55
2.2.4 實驗試劑配製 55
2.2.5 毛細管之前處理 57
2.2.6 DNA分離 57
2.3 實驗結果與討論 58
2.3.1 pH與EtBr的影響 58
2.3.2 GNPs、PEO與EtBr濃度對DNA分離的影響 61
2.3.3 分離8 bp-23.1 kbp 之DNA混合物 63
2.4 結論 64
2.5 參考文獻 66
2.6 本章圖表 69
第三章 以光合成及光沉降法在蛋白質存在下合成金奈米粒子 76
3.1 導論 76
3.2 實驗 78
3.2.1實驗試藥 78
3.2.2實驗試劑配製 78
3.2.3金奈米粒子之吸收光譜與粒徑大小測量 80
3.3 實驗結果與討論 81
3.3.1 BSA與金離子濃度對合成反應的影響 81
3.3.2 pH與溫度變化對合成反應的影響 84
3.3.3 蛋白質晶片 87
3.4 結論 88
3.5 參考文獻 90
3.6 本章圖表 93
第四章 總結 100
1.8 參考文獻34

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