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.
出处
《中成药》
CAS
CSCD
北大核心
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)
关键词
RAPD
柴胡
正交设计
综合评分法
RAPD
Bupleurum chinense DC
orthogonal design
comprehensive scoring method
