陳炳煇臺灣大學:機械工程學研究所侯建成Hou, Chien-ChengChien-ChengHou2010-06-302018-06-282010-06-302018-06-282009U0001-2807200916325900http://ntur.lib.ntu.edu.tw//handle/246246/187183The concept of Polymerase Chain Reaction primarily uses those three processes (Denaturizing, Annealing and Extension) as one cycle, and DNA will clone by the increase rate of 2n. Real-Time PCR is the most famous in the concept. The technique mainly needs to combine Thermal Control Cycler with Fluorimeter, and do DNA outside-body proliferation in thermal control process. And meanwhile, in order to monitor Labeling Dye in proliferation process, the change of fluorescence level and do quantitative analysis, the instrument called Real-time PCR machine is needed to achieve the above-mentioned technique. So Thermal Cycler and Fluorimeter are the keys to quantitative analysis.he first part of the thesis will discuss the difficulty that Lab-on-a-Chip is heated-from-the bottom system. When Heated-from-the bottom system controls the bottom heated temperature to do the Polymerase Chain Reaction process, it is possible to make inner reagent cause the phenomena of temperature gradient and natural convection. When Using thermocouple measures the bottom temperature of Lab-on-a -Chip and upper temperature of reagent, we find there is a gradient temperature up to 9.7℃.In order to solve the problem, changing reagent volume or adding DMSO to reagent are proposed.he second part of the thesis will discuss how the fluorescence detection system operate in DNA fluorescence detection. In the study, we use 350bp(base pair) Hepatitis B virus, with 108 copies/ml DNA template as experiment reagent, and besides, adding SyBr Green dye for fluorescence detection. Because fluorescence signal is invisible to the naked eye, we use MATLAB to cope with the fluorescence signals filmed by CCD to quantify them. Thus we can get weak DNA fluorescence signals which are used for quantitative analysis.目錄要 ibstract iii號說明 v文字母符號 v臘符號 vii錄 viii目錄 xi目錄 xii一章 緒論 1.1 前言 1.2 研究動機與目的 2.3 文獻回顧 2.4 論文架構 6二章 研究方法與原理 12.1 聚合酶連鎖反應(polymerase chain reaction) 12.1.1 PCR原理 12.1.2 PCR影響因素 13.2 熱傳遞分析 16.3 洋菜凝膠電泳 17.3.1 凝膠電泳原理 17.3.2 利用膠電泳觀察PCR複製的結果 19.4 控制原理 20.4.1 PWM原理 20.4.2 PID原理 20.5 即時偵測原理 21.5.1 螢光激發理論 21.5.2 DNA結合螢光探針原理 22.5.3 螢光分析儀的收光效率 23三章 實驗設備與研究步驟 29.1 底部加熱型PCR機台 29.1.1 加熱載具 29.1.2 控制器 29.2 螢光儀 31.2.1 光學機構腔體設計 32.2.2 濾光鏡(Filter) 32.2.3 激發光源 32.2.4 電荷耦合攝影機(Charge Coupled Device) 33.3 洋菜膠體電泳與紫外光檢測系統 33.3.1 洋菜膠體模具 33.3.2 電泳槽 33.3.3 紫外光檢測系統 33.4 傳統PCR機台 34.5 溫度量測系統 34.6 實驗試劑與化學藥品 34.6.1 聚合酶連鎖反應實驗試劑與化學藥品 34.6.2 洋菜膠體電泳檢測所用的試劑與化學藥品 36.6.3 拋棄式PMDS製作所用的試劑與化學藥品 37.7 研究步驟 37四章 結果與討論 59.1 平底加熱型PCR機台的問題與探討 59.1.1 改變試劑體積的實驗結果 59.1.2 試劑體積與溫度梯度探討 60.1.3 DMSO處理 62.2 螢光偵測 63.2.1 不同體積下的螢光訊號 63.2.2 即時偵測PCR實驗 64五章 結論與未來展望 80.1 結論 80.2 未來展望 80考文獻 82錄A 877840540 bytesapplication/pdfen-US聚合酶連鎖反應二甲基亞B肝病毒雷利數PCRDMSOHBVRayleigh Number[SDGs]SDG3thesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/187183/1/ntu-98-R96522317-1.pdf