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| 七叶树RAPD反应体系的优化 | |
| 引用本文： | 王绍文,时明芝,张建国,曹兴.七叶树RAPD反应体系的优化[J].辽宁林业科技,2010(2):18-20. |
| 作者姓名： | 王绍文 时明芝 张建国 曹兴 |
| 作者单位： | 1. 聊城大学农学院,山东,聊城,252059 2. 聊城大学农学院,山东,聊城,252059;山东财政学院,山东,济南,250014 3. 济宁市林业局,山东,济宁,272019 |
| 基金项目： | 山东省教育厅科技攻关资助项目(J07YG08) |
| 摘 要： | 以七叶树新鲜叶片为材料,研究了七叶树RAPD分析过程中的影响因素包括Taq酶、Mg2+、dNTP、引物、模板DNA浓度、变性时间、循环次数等,建立了适合七叶树RAPD反应的PCR体系。即20μl反应体系中含有dNTP 0.25 mmol/L,Mg2+2.5 mmol/L,Taq酶1.0U,引物0.8μmol/L,DNA模板50ng。扩增程序为:94℃预变性5min,然后40个循环(94℃变性30 s,33℃退火40 s,72℃延伸60 s),最后72℃延伸10min,4℃保存。 |
| 关 键 词： | 七叶树 RAPD 优化 |
Optimization of RAPD reaction systems for Aesculus chinensis |
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| WANG Shao-wen,SHI Ming-zhi,ZHANG Jian-guo,CAO Xing.Optimization of RAPD reaction systems for Aesculus chinensis[J].Liaoning Forestry Science and Technology,2010(2):18-20. | |
| Authors: | WANG Shao-wen SHI Ming-zhi ZHANG Jian-guo CAO Xing |
| Institution: | 1.College of Agriculture/a>;Liaocheng University of Sandong Province/a>;Liaocheng 252059/a>;China/a>;2.Shandong College of Finance and Economics/a>;Jinan 250014/a>;3.Jining City Forestry Bureau/a>;Jining 272019/a>;China |
| Abstract: | Taking flesh leaves of Aesadus chinensis as materials, such factors impacted on RAPE） analysis as Taq enzyntes, Mg2＋, dNTP, primer, concentration of DNA template, time of degeneration and cycling were studied, and PCR systems fitted for RAPE） reaction of Aeculs chinensis were established. There were 0.25mmol/L of dNTP, 2.Smmol/L of Mg2＋, 1.0U of Taq （mzyrncs, 0.81μml/L of primer and 50ng of DNA template in 20μl reaction systems. The amplification program was as follows： pr 5min at 94 μ, and then cycling for 40 times （degeneration 30s at 94 μ, annealing 40s at 33 μ extension 60s at 72 μ）, extension 10min at 72 μ at Last. |
| Keywords: | Aesculus chinensis RAPD optimization |
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