七叶树RAPD反应体系的优化  

Optimization of RAPD reaction systems for Aesculus chinensis

作　　者：王绍文[1] 时明芝[1,2] 张建国[3] 曹兴[1] 

机构地区：[1]聊城大学农学院,山东聊城252059 [2]山东财政学院,山东济南250014 [3]济宁市林业局,山东济宁272019

出　　处：《辽宁林业科技》2010年第2期18-20,共3页Liaoning Forestry Science and Technology

基　　金：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。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.

关 键 词：七叶树 RAPD 优化 

分 类 号：S793[农业科学—林木遗传育种] Q943[农业科学—林学]