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題 名 | 細胞數、蛋白酶處理與聚合酶對於SRY/ZFX與BE5引子進行性別鑑定的影響=Effects of Cell Numbers, Proteinase K Treatment and Polymerase on Sex Determination with SRY/ZFX and BE5 Primers |
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作 者 | 蕭振文; 陳裕信; 蔡麗卿; 楊鎮榮; 李善男; 鄭登貴; 薛佑玲; 陳立人; | 書刊名 | 畜產研究 |
卷 期 | 37:4 2004.12[民93.12] |
頁 次 | 頁333-341 |
分類號 | 437.21 |
關鍵詞 | 牛胚; 性別鑑定; 引子; 聚合酶連鎖反應; Bovine embryo; Sex determination; Primer; PCR; |
語 文 | 中文(Chinese) |
中文摘要 | 本研究之主要目的乃探討應用SRY/ZFX及BE5引子進行聚合酶連鎖反應(polymerase chain reaction, PCR)性別鑑定時,細胞數、PCR之前蛋白酶K(proteinase K)處理與否,以及聚合酶種類對於性別鑑定敏感性與特異性之影響,藉以提昇性別鑑定之準確性。試驗分別利用經過純化不同性別乳牛基因組DNA及體外培養之乳牛體細胞(卵丘細胞)做為性別鑑定之材料。結果顯示,經過純化之不同性別乳牛基因組DNA樣品,SRY/ZFX及BE5二組引子之性別鑑定敏感性可達到5 ng,且不論一般Taq聚合酶或於溫條件始啟動聚合作用的Hot Start聚合酶均可獲得專一性的PCR產物;而利用體外培養的乳牛體細胞進行分析時,細胞樣品至少需10個以上,並於PR前先經蛋白酶K前處理,配合使用一般Taq或Hot Start DNA聚合酶進行PCR,是適合的細胞樣品性別鑑定條件。 |
英文摘要 | The purpose of this study was to determine the effects of cell numbers, proteinase K treatment prior to polymerase chain reaction (PCR0, and polymerase on the sensitivity and specificity of sex determination by using SRY/ZFX or BE5 as primer. Purified genomic DNA from male and female dairy cattle and in-vitro-cultured cumulus cells were used for the sex determination. The results showed that the sensitivity of sex determination of genomic DNA was 5 ng when Taq or Hot Start polymerase was used in amplification with either SRY/ZFX or BES primers. When the somatic cells were used as samples for determination, treatment with proteinase K and then PCR running with Hot Start DNA polymerase, sexing efficiency would be effectively improved. Ten cells were the minimal numbers required for the successful sexing. |
本系統中英文摘要資訊取自各篇刊載內容。