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珍稀濒危植物永瓣藤DNA提取与ISSR条件优化
DNA extraction and optimization of ISSR reaction system for rare and endangered plant Monimopetalum chinense
【摘要】 以硅胶干燥叶片为材料,研究永瓣藤DNA的提取方法,并对影响简单重复序列区间分子标记反应的各条件进行了优化。建立了永瓣藤ISSR的优化反应体系和程序,即在20μL反应体系中,含20ng模板DNA、2.375mmol/L Mg2+、0.15mmol/L dNTPs、1.5U Taq DNA聚合酶、225nmol/L随机引物;扩增程序为:94℃预变性5min;然后94℃15s、48~50℃45s、72℃1min,35个循环,最后72℃延伸10min,4℃保存。本研究为进行永瓣藤种群遗传多样性研究奠定了基础。
【Abstract】 In the present study,DNA of Monimopetalum chinense was extracted from the silica gel dried leaves.The optimized reaction system and programme of inter simple sequence repeats(ISSR)for M.chinense are as follows:the reactions were performed in a volume of 20 μL containing 20 ng of template DNA,2.375 mmol/L Mg2+,0.15 mmol/L dNTPs,1.5U Taq DNA polymerase,225 nmol/L primer;thermal cycling was programmed by 1 cycle for 5 min at 94 ℃,followed by 35 cycles for 15 s at 94 ℃,45 s at 48~50 ℃ and 1 min at 72 ℃,and finally,by 1 cycle for 10 min at 72 ℃,stored at 4 ℃.
【Key words】 Monimopetalum chinense; rare and endangered plant; DNA extraction; ISSR;
- 【文献出处】 广西植物 ,Guihaia , 编辑部邮箱 ,2007年06期
- 【分类号】Q943.2
- 【被引频次】10
- 【下载频次】220