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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39174
標題: | 薄層聚合酵素連鎖反應系統與螢光檢測系統之開發 Development of a Thin Film Polymerase Chain Reaction and Fluorescence Detection System |
作者: | Guo – Cheng He 何國誠 |
指導教授: | 朱元南(Yuan-Nan Chu) |
關鍵字: | 聚合酵素連鎖反應:螢光檢測:凝膠電泳, Polymerase chain reaction (PCR):fluorescence detection: gel electrophoresis, |
出版年 : | 2005 |
學位: | 碩士 |
摘要: | 本論文研究薄層聚合酵素連鎖反應(PCR)系統及螢光檢測系統,薄層聚合酵素連鎖反應系統之反應槽以PDMS做為材質,並且以單一鑄模方式製作,製程簡單且具有高製造良率。昇降溫部份以熱電致冷晶片做為加熱元件,搭配離心風扇做強制對流泠却。溫控部份以LabVIEW軟體擷取Thermocouple量測的溫度資料,再進行PID控制以改變TEC的加熱功率輸出。經實驗證明此薄層聚合酵素連鎖系統可成功的複製DNA序列。螢光檢測系統分為二部份裝置,雷射誘導螢光檢測部份以532nm 波長的綠光半導體雷射做為激發光源,搭配適當光學元件,最後以光電倍增管偵測樣本螢光訊號,並以 LabVIEW 軟體程式做為資料擷取系統。螢光影像系統則以UV光做為DNA染料激發光源,再以CCD擷取螢光影像。本螢光檢測系統利用樣本步進平台可作定點或掃描之螢光偵測,達成多用型螢光檢測系統之目的,實驗證實可成功的偵測DNA螢光訊號。 A thin-film Polymerase Chain Reaction (TF-PCR) system and a fluorescence detection system have been developed. Made of PDMS, the TF-PCR is fabricated by casting with easy of manufacturing and very high quality. The TF-PCR contains thermoelectric cooling components (TEC) as heaters, and uses a centrifuge system fan for forced convention cooling. The temperature controlling system is composed of a thermocouple as a temperature sensor, a DAQ card gather to voltage signals and a PC with the LabVIEW program. The monitoring and controlling of the TEC heater is through a PID program. Successful amplification of DNA strains of a shrimp virus with 120 and 226 base pair has been demonstrated by this TF-PCR. The fluorescence detection system can be divided into two subsystems. The LASER-induced fluorescence detecting subsystem includes a Green color semiconductor LASER (532nm in wavelength) as an exciting light source, followed by proper optical components and a photo multiplying tube (PMT), then fluorescence signals from DNA samples can be caught. The fluorescence image subsystem has a UV light source and can extract images from a CCD camera. This fluorescence detection system also uses step motors to control the sample stand, so that fixed scanning or moved scanning is possible. This fluorescence detection system is a multiple functional detecting system. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39174 |
全文授權: | 有償授權 |
顯示於系所單位: | 生物機電工程學系 |
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