摘要
用改良CTAB法从巨龙竹硅胶干燥的叶片中提取基因组DNA,成功地进行RAPD扩增。通过正交法对RAPD反应条件优化,6个试验因子的最适条件为:模板DNA1～1.5ng/μl,引物0.8～1.0μmol/L,dNTPs0.1～0.15mmol/L,Mg^(2+) 2.0～2.5mmol/L,Taq酶0.5～1.0U,退火温度35～36℃。
An improved CTAB method of DNA extraction is applied to obtaining genomic DNA from silica gel dried leaves of Dendrocalamus sinicus Chia et J. L. Su. The DNA is successfully used for RAPD analysis. In this paper, six essential factors that might affect the result of RAPD are compared and optimized by applying orthogonal experiment. The result shows that the optimal conditions of six factors in RAPD - PCR reaction system are DNA 1 ~ 1. 5ng/μl; random primer 0.8-1.0/μol/L; dNTPs 0.1-0.15mmol/L; Mg2+ 2. 0 ~ 2. 5mmol/L; Taq polymerase 0.5- 1.0U; annealing 36-37℃.
出处
《昆明理工大学学报（理工版）》
2003年第1期127-131,共5页
Journal of Kunming University of Science and Technology(Natural Science Edition)
基金
国家教委资助项目
