利用综合评分法优化柴胡ISSR-PCR反应体系  

Optimization for ISSR -PCR Reaction System of Bupleurum chinense DC.by Comprehensive Scoring Method

作　　者：杨旻[1] 胡继鹰[1] 

机构地区：[1]湖北中医药大学,湖北武汉430065

出　　处：《现代中药研究与实践》2013年第3期23-24,22,共3页Research and Practice on Chinese Medicines

基　　金：湖北省自然科学基金(2010CDB07104)

摘　　要：目的研究柴胡ISSR-PCR的最佳反应体系。方法应用综合评分法,对Taq酶、Mg2+、引物、总DNA和dNTPs 5种ISSR反应成分进行优选。结果优化反应体系为:25μL反应体系中含有1×buffer、Taq酶1.0 U、Mg2+2.5 mmol.L-1、引物0.4μmol.L-1、DNA 75 ng、dNTPs 0.3 mmol.L-1。结论综合评分法优化柴胡ISSR反应体系可行,为下一步试验提供了可靠参数。Objective To optimize the ISSR-PCR reaction system ofBupleurum chinense DC. Methods The comprehensive scoring method was applied to evaluate the effects of different concentrations of Taq polymerase, Mg^2＋, dNTPs, primer and genomic DNA on Bupleurum chinense DC ISSR-PCR results. Results A suitable ISSR reaction system was developed, i.e. l×Taq polymerase reaction buffer, 2.5 mmol·L^-1 Mg^2＋, 1.0 U Taq DNA polymerase, 0.3 mmolo·L^-1 dNTPs, 0.4 μmol·L-1 primer and 75 ng genomic DNA templates with total 25 μL reaction solution. Conclusion The results showed that the comprehensive scoring method could be used successfully to optimize the ISSR-PCR system and provided a reliable parameter for next expeririment.

关 键 词：综合评分 ISSR 柴胡 正交设计 

分 类 号：R282[医药卫生—中药学]