摘要
以麻竹DNA为模板,对影响麻竹RAPD扩增的重要参数进行了优化试验,以期建立麻竹RAPD反应的最适体系。最终得出的麻竹RAPD反应体系为:20μL反应体系,2μL10倍反应缓冲液,模板含量为50ng,2 5mmol·L-1的Mg2+,1 5U的Taq酶,dNTP为1 75mmol·L-1,引物浓度为0 4μmol·L-1。优化后的RAPD反应程序为:94℃3min→[94℃1min→37 5℃1min→72℃1min20sec]40个循环→72℃8min→4℃保持。
Based on the genomic DNA of Dendrocalamus latiflorus ,some essential factors that might affect the results of RAPD were compared and the optimal RAPD reaction system for D.latiflorus was established.The optimized condition for 20 μL reaction system were 2 μL 10 diploid buffer, 50 ng template DNA, 2. 5 mmol·L-1 MgCl2,1. 5 U Taq DNA polymerase, 1. 75 mmol·L-1 dNTP and 0. 4 μmol·L-1 Random primer.The optimized reaction program was initially at 94 ℃ for 3 min, followed by 40 cycles at 94 ℃ for 1 min, at 37. 5 ℃ for 1 min, at 72 ℃ for 1 min 20 sec, and then held at 72 ℃ for 8 min,and finally kept at 4 ℃.
出处
《林业科学研究》
CSCD
北大核心
2003年第5期554-559,共6页
Forest Research
基金
国家林业部指南项目"竹类植物遗传多样性研究和遗传连锁图谱的构建"(项目编号190)
