| 石榴ISSR-PCR反应体系的正交设计优化 |
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| 引用本文： | 赵丽华.石榴ISSR-PCR反应体系的正交设计优化[J].北方园艺,2010(19). |
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| 作者姓名： | 赵丽华 |
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| 作者单位： | 西昌学院农业科学学院;西南大学园艺园林学院;重庆市花卉工程技术研究中心; |
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| 基金项目： | 四川省教育厅科研基金，西昌学院科研课题资助项目 |
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| 摘 要： | 现利用正交设计L16(45)探讨DNA模板、Mg2+、dNTPs、引物、TaqDNA聚合酶对石榴ISSR-PCR反应体系的影响,首次应用加权平均法及百分制对正交实验结果进行评分,并应用DPS数据处理系统进行数据分析;同时对石榴ISSR-PCR最佳反应体系进行退火温度梯度试验。结果表明:适合石榴的ISSR-PCR最优反应体系(25μL)为:Mg2+1.75 mmol/L、dNTPs0.15 mmol/L、TaqDNA聚合酶1 U、引物0.8 mmol/L、DNA模板20 ng;引物UBC 811的最适退火温度为51.2℃,且最适退火温度因引物而异。
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| 关 键 词： | 石榴 正交设计 ISSR 反应体系 优化 |
Optimization of ISSR-PCR Reaction System for Pomegranate in Orthogonal Design |
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| ZHAO Li-hua.Optimization of ISSR-PCR Reaction System for Pomegranate in Orthogonal Design[J].Northern Horticulture,2010(19). |
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| Authors: | ZHAO Li-hua |
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| Institution: | ZHAO Li-hua1,2(1.Agricultural Sciences Institute,Xichang College,Xichang,Sichuan 615013,2.College of Horticulture and Landscape Architecture,Southwest University,Flower Engineering Technology Research Center,Chongqing 400715) |
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| Abstract: | The optimize ISSR-PCR reaction system of pomegranate was studied,the orthogonal design L16(45)was used to optimize 5 factors Taq DNA polymerase,Mg2+,DNA template,dNTPs and primerof ISSR-PCR amplification system for pomegranate,the weighted average method and hundred mark system are used to evaluate the results of the orthogonal scoring,the DPS data processing systems was used data analysis;And then do annealing temperature gradient test to optimize ISSR-PCR reaction system.The results showed that the best I... |
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| Keywords: | pomegranate orthogonal design ISSR reaction system optimization |
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