| 青海云杉ISSR-PCR反应体系的建立与优化(英文) |
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| 引用本文： | 王文峰,李毅,马彦军,李栋栋.青海云杉ISSR-PCR反应体系的建立与优化(英文)[J].Agricultural Science&Technology,2009(3). |
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| 作者姓名： | 王文峰 李毅 马彦军 李栋栋 |
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| 作者单位： | 甘肃农业大学林学院; |
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| 摘 要： | Objective] The experiment aimed to determine the optimum ISSR-PCR reaction system of Picea crassifolia kom. Method] Picea crassifolia kom. was used as material to select and optimize influencing factors of ISSR-PCR such as Mg2+, dNTPs, Taq DNA polymerase, template DNA, primers, annealing temperature. Result] The optimum ISSR-PCR reaction system in 20 μl reaction system was consisted of 1 μl 10×buffer, 1.5 mmol/L Mg2+, 0.2 mmol/L dNTPs, 1.0 U Taq DNA polymerase, 40 ng template DNA, 0.6 μmol/L primers. According to gradient test of annealing temperature in optimum ISSR-PCR reaction system of Picea crassifolia kom, it was found that the optimum annealing temperature of UBC 818 was 54.2 ℃ and the annealing temperature was different for different primers.Conclusion]The construction of ISSR-PCR reaction system provided technical basis for classification of germplasm resources, construction of genetic map, gene mapping of Picea crassifolia kom. through using ISSR technology.
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| 关 键 词： | 聚合酶 反应体系 扩增产物 退火温度 引物 浓度 青海云杉 优化 |
Establishment and Optimization of ISSR-PCR Reaction System for Picea crassifolia kom. |
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| WANG Wen-feng,LI Yi,MA Yan-jun,LI Dong-dong College of Forestry Science,Gansu Agricultural University,Lanzhou.Establishment and Optimization of ISSR-PCR Reaction System for Picea crassifolia kom.[J].农业科学与技术(英文版),2009(3). |
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| Authors: | WANG Wen-feng LI Yi MA Yan-jun LI Dong-dong College of Forestry Science Gansu Agricultural University Lanzhou |
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| Institution: | WANG Wen-feng,LI Yi,MA Yan-jun,LI Dong-dong College of Forestry Science,Gansu Agricultural University,Lanzhou 730070 |
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| Abstract: | Objective] The experiment aimed to determine the optimum ISSR-PCR reaction system of Picea crassifolia kom. Method] Picea crassifolia kom. was used as material to select and optimize influencing factors of ISSR-PCR such as Mg2+, dNTPs, Taq DNA polymerase, template DNA, primers, annealing temperature. Result] The optimum ISSR-PCR reaction system in 20 μl reaction system was consisted of 1 μl 10×buffer, 1.5 mmol/L Mg2+, 0.2 mmol/L dNTPs, 1.0 U Taq DNA polymerase, 40 ng template DNA, 0.6 μmol/L primers. According to gradient test of annealing temperature in optimum ISSR-PCR reaction system of Picea crassifolia kom, it was found that the optimum annealing temperature of UBC 818 was 54.2 ℃ and the annealing temperature was different for different primers.Conclusion]The construction of ISSR-PCR reaction system provided technical basis for classification of germplasm resources, construction of genetic map, gene mapping of Picea crassifolia kom. through using ISSR technology. |
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| Keywords: | Picea crassifolia kom ISSR System establishment Optimization |
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