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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, 0(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
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, 0(2): 18-20
Authors:WANG Shao-wen  SHI Ming-zhi    ZHANG Jian-guo  CAO Xing
Affiliation: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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