| 葡萄目标起始密码子多态性反应体系的优化 |
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| 引用本文： | 张君玉,郭大龙,龚莹,刘崇怀,李猛,张国海.葡萄目标起始密码子多态性反应体系的优化[J].果树学报,2011(2):209-214. |
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| 作者姓名： | 张君玉 郭大龙 龚莹 刘崇怀 李猛 张国海 |
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| 作者单位： | 河南科技大学林学院;中国农业科学院郑州果树研究所; |
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| 基金项目： | 国家自然科学基金(NSFC:30800742); 河南省教育厅自然科学研究计划(2009B210003); 国家葡萄产业技术体系(nycytx-30-zy-01) |
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| 摘 要： | SCoT标记是一种新型目的基因分子标记方法,采用L16(45)正交试验和单因素试验2种方法优化了葡萄SCoT-PCR反应体系,结果表明正交试验中关键影响因子为dNTPs浓度,单因素试验中为dNTPs和Mg2+浓度.综合2种方法,优化的葡萄SCoT-PCR反应体系为:Mg2+2.0 mmol·L-1、dNTPs 0.3 ...
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| 关 键 词： | 葡萄 SCoT 正交设计 单因素试验 体系优化 |
Optimization of Start Codon Targeted Polymorphism PCR(SCoT-PCR) system in Vitis vinifera |
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| ZHANG Jun-yu,GUO Da-long,GONG Ying,LIU Chong-huai,LI Meng,ZHANG Guo-hai.Optimization of Start Codon Targeted Polymorphism PCR(SCoT-PCR) system in Vitis vinifera[J].Journal of Fruit Science,2011(2):209-214. |
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| Authors: | ZHANG Jun-yu GUO Da-long GONG Ying LIU Chong-huai LI Meng ZHANG Guo-hai |
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| Institution: | ZHANG Jun-yu1,GUO Da-long1,GONG Ying1,LIU Chong-huai2,LI Meng1,ZHANG Guo-hai1 (1College of Forestry,Henan University of Science & Technology,Luoyang,Henan 471003 China,2Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou,Henan 450009 China) |
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| Abstract: | The grape SCoT-PCR reaction system was optimized based on the L16(45) orthogonal experiment and single factor experiment design.The results showed that: the key factor was the dNTPs concentration in the orthogonal experiment,while in the single factor experiment,the key factors were the dNTPs and Mg2+ concentration;Based on the results,it was suggested that the optimal SCoT-PCR system for grape was: total 20 μL reaction system containing 2.0 mmol·L-1 Mg2+,0.3 mmol·L-1 dNTPs,1.0 U Taq polymerase,0.3 μmol·L-1 primer,and 30 ng DNA template.The PCR amplification profile was pre-denaturing at 94 ℃ for 5 min,,denaturing at 94 ℃ for 1 min,annealing at 54 ℃ for 1 min and extending at 72 ℃ for 2 min,34 cycles and extending at 72 ℃ for 8 min finally.The optimal reaction system was further verified in 20 grape varieties,which showed good stability and repeatability.The system provides a new technology for evaluation of genetic diversity,genomic analysis,construction of fingerprinting and molecular marker assisted breeding for the grape. |
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| Keywords: | Grape(Vitis vinifera) SCoT Orthogonal design Single factor experiment System optimization |
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