| 多重PCR鉴定水牛胚胎性别的研究 |
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| 引用本文： | 付强,郑海英,陈焕华,张明,卢克焕. 多重PCR鉴定水牛胚胎性别的研究[J]. 广西农业科学, 2011, 0(8): 984-986 |
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| 作者姓名： | 付强 郑海英 陈焕华 张明 卢克焕 |
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| 作者单位： | 亚热带农业生物资源保护与利用国家重点实验室;中国科学院广西水牛研究所; |
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| 基金项目： | 广西青年科学基金项目(2011GXNSFB018028); 广西自然科学基金项目(桂科自0991047) |
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| 摘 要： | 【目的】建立多重PCR鉴定水牛胚胎性别的技术体系,为下一步优化多重巢式PCR方法奠定基础。【方法】采用热裂解法、蛋白酶K＋二硫苏糖醇（DTT）法和吐温-20＋DTT法分别提取水牛卵裂球DNA,比较3种方法对G3PDH基因的扩增效果;同时设计1对检测引物和1对内标引物用于胚胎性别鉴定,提取胚胎DNA后进行多重PCR检测。【结果】吐温-20＋DTT法具有良好的裂解效果,其有效检出率为97.1%;而经吐温-20＋DTT法抽提的DNA用于多重PCR检测,其胚胎鉴定率达到100.0%。【结论】采用吐温-20＋DTT法处理水牛胚胎样品后再进行多重PCR性别鉴定可以简化操作步骤、提高鉴定效率,但样品数量不宜太少。
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| 关 键 词： | 水牛胚胎 性别鉴定 多重PCR |
Sex identification of buffalo embryo using multiple-PCR |
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| FU Qiang,ZHENG Hai-ying ,CHEN Huan-hua,ZHANG Ming ,LU Ke-huan. Sex identification of buffalo embryo using multiple-PCR[J]. Guangxi Agricultural Sciences, 2011, 0(8): 984-986 |
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| Authors: | FU Qiang ZHENG Hai-ying CHEN Huan-hua ZHANG Ming LU Ke-huan |
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| Affiliation: | FU Qiang1,ZHENG Hai-ying 2,CHEN Huan-hua1,ZHANG Ming1 *,LU Ke-huan1 (1 State Key Laboratory for Conservation and Utilization of Subtropical Agriculture Bioresources,Nanning 530005,China,2 Guangxi Buffalo Research Institute,Chinese Academy of Agricultural Sciences,Nanning 530001,China) |
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| Abstract: | [Objective]The present experiment was conducted to develop a fast and accurate PCR based method for sex identification of buffalo embryos by PCR for providing the basis of optimizing multiple nested PCR method. [Method]Three methods to extract DNA from buffalo blastomere, including thermal lyses, proteinase K+DTT and DTT+Tween-20, were used and the isolated DNAs were comparatively analyzed for their amplification efficiency using primers based on G3PDH gene of buffalo. The embryonic DNA was amplified by usi... |
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| Keywords: | buffalo embryo sex identification multiple-PCR |
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