| 大字杜鹃RAPD反应体系的优化 |
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| 引用本文： | 姬文秀,金东淳,李虎林,鲁艺.大字杜鹃RAPD反应体系的优化[J].延边大学农学学报,2009,31(1):21-25. |
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| 作者姓名： | 姬文秀 金东淳 李虎林 鲁艺 |
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| 作者单位： | 延边大学农学院农学系,吉林,龙井,133400 |
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| 摘 要： | 以大字杜鹃为材料，以改进的CTAB法提取大字杜鹃叶片总DNA，分别就模板DNA浓度，引物浓度，DNTP浓度，TaqDNA聚合酶量及镁离子浓度对大字杜鹃RAPD反应结果的影响进行研究．通过对各因子的组合比较，建立了大字杜鹃RAPD反应的优化体系，即总反应体积为25pL，其中包括10×Taq酶配套缓冲液2．5pL，模板DNA浓度50mg／L，引物浓度0．5μmol／L，dNTP浓度0．15mmol／L，TaqDNA聚合酶1U，Mg^2＋浓度2．0mmol／L，其余用重蒸馏水补足．扩增程序为94℃预变性5min，94℃变性1min，38℃退火40S，72℃延伸2min，共41个循环，72℃延伸7min．
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| 关 键 词： | 大字杜鹃 RAPD 反应体系 优化 |
Optimization on RAPD reaction system of Rhododendron schlippenbachin Maxin |
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| JI Wen-xiu,JIN Dong-chun,LI Hu-lin,LU Yi.Optimization on RAPD reaction system of Rhododendron schlippenbachin Maxin[J].Journal of Agricultural Science Yanbian University,2009,31(1):21-25. |
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| Authors: | JI Wen-xiu JIN Dong-chun LI Hu-lin LU Yi |
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| Institution: | (Agricultural Department , Agricultural College of Yanbian University ,Longjing Jiln 133400,China) |
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| Abstract: | The genomic DNA was extracted from Rhododendron schlippenbachin Maxim . by means of advanced CTAB method. Studied the influences of the concentration of template DNA, primer, dNTP, TaqDNA polymerase and Mg^2＋ on RAPD analysis procedure. Astablished the optimization system of RAPD on the base of the comparation among various influence factor groups：total volume 25μL, comprising of 10 × Taq buffer（2.5 μL）, template DNA concentration 50 mg/L, primer concentration 0. 5 μmol/L, dNTP 0. 15 mmol/L, TaqDNA polymerase 1 U, Mg^2＋concentration 2.0 mmol/L and the supplementary ddH20. The extended program was： 94℃ predenaturation 5 rain, 94 ℃denaturation 1 rain, 38℃annealing 40 s, 72℃ extension 2 rain, 41 cycles in all, 72 ℃ extension 7 min finally. |
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| Keywords: | RAPD |
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