| 应用MGB荧光探针技术确定猪KIT基因拷贝重复数 |
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| 引用本文： | 王韬,唐辉,曾勇庆,李晓宁,闫超,董林松,于希江.应用MGB荧光探针技术确定猪KIT基因拷贝重复数[J].畜牧兽医学报,2011,42(3). |
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| 作者姓名： | 王韬 唐辉 曾勇庆 李晓宁 闫超 董林松 于希江 |
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| 作者单位： | 山东农业大学动物科技学院,泰安,271018 |
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| 基金项目： | 山东省中青年科学家奖励基金(2008BS07009); 山东省科技攻关(2009GG10009036); 国家863重大专项(2008101002) |
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| 摘 要： | 旨在建立一种简便、快速检测猪KIT基因拷贝重复数(CNVs)的方法并应用于未知样品的检测。在KIT外显子2中设计TaqMan-MGB探针和引物,建立荧光实时定量PCR检测KIT基因CNVs的方法。结果表明,TaqMan-MGB探针扩增KIT基因目的片段,不同DNA梯度浓度Ct值差异极显著(P<0.001),变异系数低(0.12%～0.26%);KIT与内标基因ESR的扩增效率相差只有2.4%;采用聚类分析推测了待测的50枚样品KIT的CNVs分别归属于2、3、4、5、6个拷贝。本研究建立的TaqMan-MGB探针实时荧光定量PCR检测KIT基因CNVs的方法,具有简便、快速、特异性和稳定性好的特点,适合于普通实验室应用。
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| 关 键 词： | 猪 显性/KIT位点 拷贝数 MGB探针 荧光定量PCR |
Detecting Copy Number Variations in Porcine KIT by TaqMan-MGB Probe |
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| WANG Tao,TANG Hui,ZENG Yong-qing,LI Xiao-ning,YAN Chao,DONG Lin-song,YU Xi-jiang.Detecting Copy Number Variations in Porcine KIT by TaqMan-MGB Probe[J].Acta Veterinaria et Zootechnica Sinica,2011,42(3). |
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| Authors: | WANG Tao TANG Hui ZENG Yong-qing LI Xiao-ning YAN Chao DONG Lin-song YU Xi-jiang |
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| Abstract: | The main objective of the present study was to establish a simple,rapid method for detecting copy number variations(CNVs) in porcine KIT gene.TaqMan-MGB probes and primers were designed according to sequence of exon 2 of KIT gene,and a real-time fluorescence quantitative PCR procedure was established for the CNVs detection.The results showed that amplification curves of KIT obtained by TaqMan-MGB probe were a series of parallel curves corresponding to 2-fold serial dilutions of DNA samples,Ct values between... |
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| Keywords: | swine dominant/KIT locus copy number MGB probe FQ-PCR |
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