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目的:优化太行菊ISSR-PCR反应体系,为利用ISSR标记进行太行菊遗传多样性研究服务。方法:采用均匀设计法优化太行菊ISSR-PCR反应体系。结果:在10μL ISSR-PCR反应体系中,模板DNA 1.2μL;引物0.8μL;2×Taq MasterMix 4.6μL。反应程序为94℃预变性5min;94℃变性1min,50℃退火1min,72℃延伸1.5min,40个循环;72℃延伸10min。结论:此反应条件适合于太行菊的ISSR-PCR反应体系,为利用ISSR标记技术研究太行菊遗传多样性奠定了基础。
OBJECTIVE: To optimize the ISSR-PCR reaction system of Taihang chrysanthemum for the study on the genetic diversity of Taihang chrysanthemum using ISSR markers. Methods: The ISSR-PCR reaction system was optimized by uniform design method. Results: In 10 μL ISSR-PCR reaction system, 1.2 μL of template DNA, 0.8 μL of primer and 4.6 μL of 2 × Taq Master Mix were used. The reaction procedure was denaturation at 94 ° C for 5 min, denaturation at 94 ° C for 1 min, annealing at 50 ° C for 1 min, extension at 72 ° C for 1.5 min for 40 cycles and extension at 72 ° C for 10 min. Conclusion: The reaction conditions are suitable for ISSR-PCR reaction system of Taihang chrysanthemum, which lays the foundation for the study of genetic diversity of Taihang chrysanthemum with ISSR marker technology.