摘要
以辣椒DNA为模板，对影响辣椒RAPD扩增的重要参数进行了优化试验，以期建立辣椒RAPD反应的最佳体系。通过采用正交试验设计的方法，对辣椒RAPD条件进行了优化，结果表明：最佳的辣椒RAPD的反应体系（15μL）中含有1×buffer，MgCl22．67mM，dNTPs0．13mM，Primer0．5μmol／L，Taq0．75U／μL，DNA40-50ng。适宜扩增条件为94℃预变性4min，再进入40个PCR循环94℃变性1min，37℃退火45s，72℃延伸1min，72℃延伸10min,4℃保存。
In the study several important parameters influencing RAPD of local Capsicum annuurn L. varieties were studied in order to establish the optimum RAPD system of local Capsicum annuum L. varieties. The RAPD reaction system was established with orthogonal design. The results showed the optimum volume of 15μL RAPD system consisted of 1 × buffer,2. 67 mmol/L MgCl2, 0. 13 mmol/L dNTPs, 0. 5 tLmol/L primer, 0. 75 u Taq DNA polymerase and 40-50 ng template DNA. The suitale PCR procedure is one cycle denaturing at 94℃ for 4min , 40 cysles which involves denaturing at 94℃ for 1 min, annealing at 37℃ for 45s and extending at 72℃ for 1 rain ,one cycle extending at 72℃ for 10 min, and then kept at 4℃.
出处
《北方园艺》
CAS
北大核心
2007年第9期7-9,共3页
Northern Horticulture
基金
怀化市科技局科技基础条件平台建设资助项目(06-6)
关键词
地方辣椒RAPD反应体系
正交设计
Local Capsicum annuum L. varieties
RAPD reaction system
Orthogonal design
