综合评分法优化柴胡RAPD-PCR反应体系  

Optimization for RAPD-PCR reaction system of Bupleurum chinense DC. by comprehensive scoring method

作　　者：杨旻[1] 胡继鹰[1] 陈科力[1] 白杨[1] 

机构地区：[1]湖北中医学院,湖北武汉430061

出　　处：《中成药》2008年第5期722-726,共5页Chinese Traditional Patent Medicine

基　　金：湖北省自然科学基金项目资助(2005ABA187)

摘　　要：目的:采用综合评分法优化柴胡RAPD-PCR的反应体系。方法:以柴胡基因组DNA为模板,应用L16(45)正交表。研究了Taq酶、Mg2+、随机引物、dNTPs和模板DNA5种RAPD反应组分浓度变化对扩增结果的影响。结果:优化反应体系为:25μL反应体系中含有1×buffer、Taq酶1.5 U、Mg2+2.5 mmol/L、引物0.4μmol/L、dNTPs 0.15 mmol/L、模板DNA50 ng。结论:综合评分法能客观高效的优化柴胡RAPD反应体系。To optimize the random amplified polymorphic DNA-Polymerase chain reaction （RAPD- PCR） system of Bupleurum chinense DC by comprehensive scoring method. METHODS： The L16（4s） orthogonal diagram was applied to studying the effects of different concentrations of Taq polymerase, Mg^2＋ , dNTPs, primer and DNA on Bupleurum chinense DC. RAPD-PCR results and to get a suitable PCR system. RESULTS： A suitable RAPD reaction system was developed, i.e. 1 × Taq polymerase reaction buffer, 2.5 mmol/L Mg^2＋ , 1.5 U Taq DNA polymerase, 0. 15 mmol/L dNTPs, 0.4 μmol/L primer and 25 ng genomic DNA templates in total 25 p,L reaction solution. CONCLUSION： The results show that the comprehensive scoring method could be used successfully to optimize the RAPD-PCR system.

关 键 词：RAPD 柴胡 正交设计 综合评分法 

分 类 号：R944.27[医药卫生—药剂学]