【摘要】 在利用ISSR技术分析齿裂菌属和皮下盘菌属遗传多样性的研究中,为获得条带清晰、重复性好的ISSR扩增结果,对影响ISSR-PCR的条件进行了筛选,确定了此类菌物ISSR-PCR反应的最适宜条件:在15μLPCR反应体系中,10倍Taq酶缓冲液1.5μL,DNA模板8ng/μL,MgCl22.5mmol/L,dNTP0.15mmol/L,引物浓度0.4μmol/L,Taq酶1.00U,ddH2O9.0μL。最佳退火温度因不同的引物而定,最佳循环次数为35次。更多还原

【Abstract】 The factors which affect the ISSR analysis in the study of the genetic diversity of Coccomyces De Not.and Hypoderma De Not.,such as concentrations of template DNA,Taq DNA polymerase,Mg2+,primer and dNTP,annealing temperature,cycling times,etc.were filtrated.The optimal ISSR-PCR conditions in the experiments were as the following:10×Taq buffer 1.5 μL,template DNA 8 ng/μL,MgCl2 2.5 mmol/L,dNTP 0.15 mmol/L,primer concentration 0.4 mmol/L,Taq DNA polymerase 1.00 U,and ddH2O 9.0 μL in total 15 μL reaction volumes.Optimal annealing temperature is decided by different primers.Optimal circular time is thirty-five.The amplification system can give clear and reproducible banding patterns.更多还原