【摘要】 以杨梅DNA为模板,对影响杨梅RAPD-PCR扩增的重要参数进行了优化试验,以期建立杨梅RAPD反应的最佳体系。通过采用正交试验设计的方法,对杨梅RAPD-PCR条件进行了优化,结果表明最佳的杨梅RAPD-PCR的反应体系(20μl)中含有1×buffer,1.0U TaqDNA聚合酶,3.0mmol/L MgCl2,0.30mmol/L dNTPs,1.5μmol/L引物和模板DNA 30-40ng。适宜的扩增条件为94℃预变性3min,再进入38个PCR循环(94℃变性30s,38℃退火30s,72℃延伸90s),72℃延伸7min,4℃保存。更多还原

【Abstract】 Regard DNA of Myrica rubra as the template,several important parameters influencing RAPD of Myrica rubra were studied in order to establish the optimum RAPD system of Myrica rubra.The optimal RAPD-PCR system was established with orthogonal design.The results showed that a total volume of 20μl RAPD-PCR system consisted of 1×buffer,1.0u Taq DNA polymerase,3.0mmol/L MgCl₂,0.30mmol/L dNTPs,1.5μmol/L primer and 30-40ng template DNA.The suitable PCR procedure is one cycle denaturing at 94℃ for 3min,38 cycles which involves denaturing at 94℃ for 30s,annealing at 38℃ for 30s and extending at 72℃ for 90s,one cycle extending at 72℃ for 7min,and then kept at 4℃.更多还原