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年度 2014
計畫名稱_中文 基因轉殖棉花、番茄、小麥檢測技術之研究
計畫名稱_英文 Studies of transgenic cotton, tomato and wheat detection echnology
計畫編號_中文 103農科-6.2.3-種-X2
計畫編號_英文 103AS -6.2.3-SS-X2
主辦單位_中文 生物技術課
主辦單位_英文 Biotechnology Section
主辦人_中文 張惠如
主辦人_英文 Hui-Ju Chang
中文摘要 為建立轉殖基因棉花的檢測作業標準流程,先進行不同核酸萃取方法及棉花種子萃取前處裡的試驗,依結果並考量檢測流程的效率與成本,採用先利用均質機將棉花種子處理15秒後,再將種殼與種仁一起以自動萃取方式來進行棉花種子樣品之核酸萃取方法。透過此方法獲得的DNA溶液以歐盟公告之檢測方法進行轉殖品項PCR定性檢測試驗,結果顯示在不同轉殖品項LLCOTTON 25、GHB614、GHB119、T304-40中,可各自得到預期79 bp、119 bp、90 bp、78 bp大小的目標片段。經過多次確認試驗後,完成建立檢測轉殖品項LLCOTTON 25及GHB614之標準作業流程。
英文摘要 In this study, we used different methods for transgenic cotton seed DNA extraction. According to DNA quality and considered the efficiency and cost of testing process, we selected the homogenizer to pretreatment 15 seconds then extracted DNA from cotton seed by automated method (Smart LabAsist-16). DNA solution is obtained by this method and used as template DNA in PCR reaction for qualitative detection. The PCR condition was follow the European Union reference reports. Results are displayed in different specific transgenic event consist of LLCOTTON 25, GHB614,GHB119, T304-40, each of them can be get the expected target fragment size as 79 bp, 119 bp, 90 bp, 78 bp. After several confirm tests, we established the standard operating procedures for the quality testing of cotton LLCOTTON 25 and GHB614