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題名 | SRY/ZFX引子進行乳牛性別鑑定之聚合酶連鎖反應=Optimization of PCR Parameters for Dairy Cattle Sexing Using SRY/ZFX Primers |
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作者 | 蕭振文; 蔡麗卿; 陳裕信; 曲鳳翔; 楊鎮榮; 陳立人; Shiau, Jen-wen; Tsai, Lee-ching; Chen, Yu-hsin; Chu, Feng-hsiang; Yang, Jenn-rong; Chen, Lih-ren; |
期刊 | 畜產研究 |
出版日期 | 20050300 |
卷期 | 38:1 2005.03[民94.03] |
頁次 | 頁57-64 |
分類號 | 437.21 |
語文 | chi |
關鍵詞 | 乳牛; 性別鑑定; 引子; 聚合酶連鎖反應; Dairy cattle; Sex determination; Primer; PCR; |
中文摘要 | 本研究旨在利用聚合酶連鎖反應(polymerase chain reaction, PCR),測試SRY/ZFX引子進行乳牛性別鑑定的最適條件。探討之PCR條件包括粘合溫度、反應體積、循環數、MgCl2濃度、聚合酶沔度與dNTP濃度等,藉以推薦適合的PCR條件,改善細胞或胚性別鑑定的效率與敏感性。試驗利用體外培養(in viro culture)之荷蘭母牛卵丘細胞或公牛之耳朵成纖維細胞做為性別鑑定之樣品,每組細胞數至少10個,先經蛋白酶K(proteinase K)前處理,再測試各種影響PCR反應之重要條件。試驗結果顯示,利用SRY/ZFX引子進行細胞樣品之性別鑑定,PCR之粘合(annealing)溫度在55~56°C、40次循環、反應體積20μI、2.5 mM之MgCl2、2~3u的Taq DNA聚合酶、200~300μM之dNTP,可獲得最佳的PCR性別鑑定結果,此最適的PCR條件將可供體細胞或田間進行牛胚性別鑑定之參考。 |
英文摘要 | The purpose of this study was to determine the suitable conditions of polymerase chain reaction (PCR) for sex determination with SRY/ZFX primers. The effects of annealing temperature, reaction volume, cyclenumber, the concentration of Taq DNA polymerase, dNTP, and MgCl, on the sensitivity and specificity of PCR sexing were determined. Samples of in vitro culture somatic cells from male and female dairy cattle were used for sex determination. The results showed that the PCR conditions consisted of annealing temperature between 55~56°C, 40 cycle number, 20μl reaction volume, 2.5 mM MgCl2, 200~300μM dNTP, and 2.5~3 u Taq DNA polymerase were most suitable for sex determination of bovine somatic cells with SRY/ZFX primers. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。