| 荧光定量PCR技术优化及在草莓上的应用 |
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| 引用本文： | 张卿,邢宇,曹庆芹,秦岭.荧光定量PCR技术优化及在草莓上的应用[J].北京农学院学报,2016,31(4):21-25. |
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| 作者姓名： | 张卿 邢宇 曹庆芹 秦岭 |
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| 作者单位： | 北京农学院农业应用新技术北京市重点实验室,北京,102206;北京农学院农业应用新技术北京市重点实验室,北京,102206;北京农学院农业应用新技术北京市重点实验室,北京,102206;北京农学院农业应用新技术北京市重点实验室,北京,102206 |
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| 基金项目： | 2016年度北京市教委科研计划一般项目，北京市属高等学校创新团队建设与教师职业发展计划项目(IDHT20140509) |
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| 摘 要： | 目的]为优化荧光定量PCR技术体系并在草莓研究中应用.方法]以二倍体草莓(Fragaria vesca‘Ruegen’)为试材,草莓当中两个β-肌动蛋白基因家族成员Actin1和Actin2为内参基因,对比分析10 μL和20 μL反应体系条件下荧光定量PCR扩增反应.结果]内参基因引物组合Actin1的扩增效率等指标优于Actin2;10μL反应体系中的扩增效率等指标优于20 μL反应体系;内参基因Actin1在10 μL反应体系下是草莓最优的荧光定量PCR技术体系.结论]优化了草莓荧光定量PCR体系并应用该技术体系检测了草莓CrRLK1Ls家族成员的时空表达情况.
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| 关 键 词： | 荧光定量PCR 内参基因 草莓 扩增效率 CrRLK1Ls |
Application of optimized fluorescence quantitative PCR technology for the research of strawberry |
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| ZHANG Qing,XING Yu,CAO Qingqin,QIN Ling.Application of optimized fluorescence quantitative PCR technology for the research of strawberry[J].Journal of Beijing Agricultural College,2016,31(4):21-25. |
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| Authors: | ZHANG Qing XING Yu CAO Qingqin QIN Ling |
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| Abstract: | Objective] In order to establish and optimize quantitative real-time PCR system for studying gene expression of strawberry.Methods] Two β-actin genes (Actin 1 and Actin2) in strawberry (Fragaria vesca ‘Ruegen’) were used reference genes in amplification reaction.The amplification efficiency and standard curve were studied in 10 μL and 20 μL reaction system conditions.Results] The results showed that the amplification efficiency and standard curve related index of Actin 1 were better than that of Actin2.The amplification efficiency and standard curve related index in 10 μL reaction system were better than that of 20μL reaction system.It is optimal quantitative real-time PCR reaction system for strawberry that Actin 1 as reference gene in 10 μL reaction system.The optimal quantitative real-time PCR reaction system was used to determine the spatiotemporal expression of CrRLK1Ls subfamily in strawberry.Conclusion] The fluorescence quantitative PCR technology was optimized and used to studied spatiotemporal expression of Strawberry.The CrRLK 1Ls subfamily may play the significant role in regulating strawberry fruit development. |
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| Keywords: | real-time PCR reference genes strawberry amplification efficiency CrRLK1Ls |
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