【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,MgCl22.67 mM,dNTPs 0.13mM,Primer 0.5μmol/L,Taq 0.75 U/μL,DNA 40~50 ng。适宜扩增条件为94℃预变性4 min,再进入40个PCR循环94℃变性1 min,37℃退火45 s,72℃延伸1 min,72℃延伸10 min,4℃保存。更多还原

【Abstract】 In the study several important parameters influencing RAPD of local Capsicum annuum 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 15 μL RAPD system consisted of 1×buffer,2.67 mmol/L MgCl2,0.13 mmol/L dNTPs,0.5 μmol/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 min,one cycle extending at 72℃ for 10 min,and then kept at 4℃.更多还原