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頁籤選單縮合
題名 | Study of PCR Detection Methods for Genetically Modified Soybeans with Reference Molecules=使用分子參考物質於Real-time PCR檢測基因改造大豆定量之研究 |
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作者姓名(中文) | 林旭陽; 魏秀安; 林富邦; 施養志; | 書刊名 | 藥物食品分析 |
卷期 | 14:2 2006.06[民95.06] |
頁次 | 頁194-202+221 |
分類號 | 412.37 |
關鍵詞 | 即時同步定量PCR; 分子參考物質; 基因改造大豆; 定量分析; Real-time PCR; PCR; ELISA; Reference molecules; GMO; Soybeans; Quantification; |
語文 | 英文(English) |
中文摘要 | 本實驗使用標準參考物質與自行研發之分子參考物質對基因改造大豆 (roundup ready soybeans,RRS) 之檢測於LightCycler real-time PCR系統試驗探討。以基因改造大豆5%、2%及1%參考物質之檢測,分別自行重被獨立試驗17、11及11次結果平均值與標準差 (mean ± SD) 分別為4.89 ± 0.45、1.98 ± 0.63與1.09 ± 0.05;比較標準參考物質與分子參考物質之適用性重複6次檢測結果,使用標準參考物質之平均值與標準差分別為5.45 ± 0.32、2.42 ± 0.13、1.20 ± 0.09,以pSAM2分子標準物質檢測結果平均值與標準差則分別為4.83 ± 0.45、2.09 ± 0.12、1.08 ± 0.10。並以ELISA、PCR與real-time PCR方法分別檢測自製豆漿於加熱過程中基因改造蛋白與殖入基因之變化,以ELISA檢測自製豆漿於加熱過程基因改造蛋白之變化,顯示在加熱75v1-3分鐘OD450值降低至最低值即無法正確檢出RRS之專性蛋白。使用real-time PCR方法,發現目標基因在100v已大多降解,但仍可萃取及檢測,而以121v加熱10分鐘以上,目標基因大幅降解,萃取檢測較為困難。 |
英文摘要 | The study of detection methods for genetically modified (GM) soy using certified reference material (CRM) and novel references molecules was operated on LightCycler real-time PCR machines system. The test results of this study demonstrated the methods used to be applicable to the specific quantitation of one line of GM soy. Independent repeat tests for 5, 2 and 1% CRM were 17, 11 and 11, and the test results mean ± SD were 4.89 ± 0.45, 1.98 ± 0.63 and 1.09 ± 0.05, respectively. Series 6 repeat test by use of CRM and references molecules, the results of CRM test were 5.45 ± 0.32, 2.42 ± 0.13 and 1.20 ± 0.09 (mean ± SD), and 4.83 ± 0.45, 2.09 ± 0.12 and 1.08 ± 0.10 for references molecules testing, to 5, 2, and 1% GM content, respectively. Further, soymilk was not detected by the ELISA method at OD450 when boiled under 70°C, 1~3 min but high to 100°C was detected by PCR method. Over 10 min under 121°C, DNA highly degradation detection was more difficult. Testing results should help support the practical detection for the GM soy. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。