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研究生:陳旺緯
研究生(外文):Wang-Wei Chen
論文名稱:TSA-FISH(TyramideSignalAmplification-FluorescenceinsituHybridization)法應用於翡翠水庫:方法建立與評估
論文名稱(外文):TSA-FISH (Tyramide Signal Amplification - Fluorescence in situ Hybridization) method applied in Fei-Tsui reservoir:Method development and assessment
指導教授:夏復國夏復國引用關係
指導教授(外文):Fuh-Kwo Shiah
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:海洋環境化學與生態研究所
學門:工程學門
學類:環境工程學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:66
中文關鍵詞:TSA-FISH法探針嚴謹度
外文關鍵詞:TSA-FISHStringency
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  • 下載下載:23
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近年來TSA-FISH (Tyramide Signal Amplification – Fluorescence in situ Hybridization)法廣泛應用於環境中微生物的研究且能有效改善傳統FISH法中螢光訊號強度過低的問題。本實驗引用TSA-FISH法進行翡翠水庫細菌群聚結構的觀測研究,主要以建立實驗方法為目標,並藉由探針嚴謹度(Stringency)測試,找出在最適formamide (FA)濃度的雜合條件下之螢光訊號強度與相對數量百分比(目標細菌數量與整體細菌數量之比例)的結果,進行方法適用性的評估。研究結果顯示,選用EUB338mix探針進行偵測時,在相對數量百分比結果上有低估現象,認為其共通性(universal)在翡翠水庫研究上仍是不足的。偵測Actinobacteria細菌群的探針HGC69a的最適FA濃度為55 % (v/v);加入競爭型探針(competitor)的最適FA濃度為50 %。另一偵測相同細菌群探針HGC236最適FA濃度為50 ~ 60 %;加入輔助型探針(helper)最適濃度也為50 ~ 60 %。比較兩者,發現在相對數量百分比上,前者得到結果較佳,因此在研究Actinobacteria細菌群時可採用HGC69a探針。其他探針測試結果,雖可得知最適FA濃度,但因觀察到目標細菌數量過低,可能潛在地影響最適FA濃度的判別,必須再加以驗證。
中文摘要........................................................................................................i
英文摘要.......................................................................................................ii
誌謝..............................................................................................................iii
目錄..............................................................................................................iv
表目錄...........................................................................................................v
圖目錄...........................................................................................................v
附錄..............................................................................................................vi

前言...............................................................................................................1
研究方法.......................................................................................................4
結果...............................................................................................................9
討論與建議.................................................................................................13
結論.............................................................................................................20
參考文獻.....................................................................................................21
表列.............................................................................................................31
圖列.............................................................................................................32
附錄列.........................................................................................................44
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