【摘要】 利用改进的CTAB法提取百里香总DNA,同时采用正交试验法对百里香基因组DNA的RAPD-PCR和ISSR-PCR反应体系及扩增条件进行优化,确定了两种标记技术的最佳PCR体系分别为:20μl RAPD反应体系中,含2.0μL 10×PCR Buffer、1.0mmol/L Mg2+、250μmol/L dNTP、1.5U Taq酶、0.8μmol/L引物和80ng模板DNA。20μl的ISSR-PCR反应体系中,含2μL10×PCR Buffer、2.0mmol/L Mg2+、250μmol/L dNTP、1U Taq酶、1.0μmol/L引物和20ng模板DNA。百里香RAPD-PCR和ISSR-PCR反应体系的建立,为今后百里香属植物的系统分类和遗传多样性分析提供一个标准化程序。更多还原

【Abstract】 An improved CTAB method of DNA extraction was applied to obtain total DNA from the fresh leaves of Thymus mongolicus Ronn.In this paper,we used orthogonal experimental method to optimize the RAPD-PCR and ISSR-PCR reaction systems.The result showed that the optimal conditions in 20μL RAPD-PCR system were 10×Buffer Mg2+ free 2μL,1.0mmol/L Mg2+,250μmol/L dNTP,1.5UTaq polymerase,0.8μmol/L random primer,80ng DNA and the optimal conditions in 20μL ISSR-PCR system were 10×Buffer Mg2+free2μL,2.0mmol/L Mg2+,250μmol/L dNTP,1UTaq polymerase,1μmol/L primer,20ng DNA.The result provided a standardizing program for systematic and analysis of genetic diversity of species in T.mongolicus Linn.更多还原