| 珍稀濒危竹种筇竹(Qiongzhuea tumidinoda)RAPD反应条件的优化 |
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| 引用本文： | 王波,高素萍,陈其兵.珍稀濒危竹种筇竹(Qiongzhuea tumidinoda)RAPD反应条件的优化[J].四川农业大学学报,2006,24(3):360-363. |
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| 作者姓名： | 王波 高素萍 陈其兵 |
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| 作者单位： | 四川农业大学,林学园艺学院,四川,雅安,625014 |
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| 基金项目： | 四川省教育厅项目(2004A030) |
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| 摘 要： | 以宜宾筇竹为材料建立了筇竹RAPD反应优化体系,用于筇竹遗传多样性分析,以改进的CTAB法提取筇竹叶片总DNA,分别测试了镁离子浓度、dNTP浓度、引物浓度、模板DNA含量、DNA聚合酶量对反应结果的影响。通过对各因子的组合比较,建立了筇竹RAPD反应优化体系:20μL PCR反应体积,10×Taq酶配套缓冲液(2μL);1.25 U Taq酶;75 ng模板DNA;45 ng引物;1.88 mmol/L MgCl2;0.19 mmol/L dNTP。
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| 关 键 词： | 筇竹 遗传多样性 优化 CTAB RAPD |
| 文章编号： | 1000-2650（2006）03-0360-04 |
| 收稿时间： | 2006-03-22 |
| 修稿时间： | 2006年3月22日 |
Optimization of RAPD Conditions of a Rare and Endangered Bamboo of Qiongzhuea tumidinoda |
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| WANG Bo,GAO Su-ping,CHEN Qi-bing.Optimization of RAPD Conditions of a Rare and Endangered Bamboo of Qiongzhuea tumidinoda[J].Journal of Sichuan Agricultural University,2006,24(3):360-363. |
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| Authors: | WANG Bo GAO Su-ping CHEN Qi-bing |
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| Abstract: | The Qiongzhuea tumidinoda RAPD reaction system optimized with yibin is established in ordered to be used in Qiongzhuea tumidinoda hereditary variety analysis.The genomic DNA of Qiongzhuea tumidinoda is extracted with improved CTAB method.The effect of content of Mg~(2+),dNTP,DNA templates,primers and DNA polymerase on experimental results is tested and the optimal reaction system of RAPD Qiongzhuea tumidinoda is determined as the following: 10×Taq polymerase corresponding buffer(2 μL),1.25 U Taq polymerase,75 ng template DNA,45 ng primer,1.88 mmol/L MgCl_2,0.19 mmol/L dNTP in total 20 μL reaction volume.This is to use RAPD marker to settle solid foundation for the classification of Qiongzhuea tumidinoda again on the molecular level. |
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| Keywords: | Qiongzhuea tumidinoda hereditary variety analysis optimization CTAB RAPD |
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