摘要
目的建立何首乌及其混伪品序列相关扩增多态性(SRAP)分子鉴定技术体系。方法正交优化影响SRAP-PCR的Mg2+、dNTPs、引物浓度与Taq聚合酶量。结果20μl反应体系的优化组合为:Mg2+2.5 mmol/L,dNTPs 0.4 mmol/L,引物0.35μmol/L,Taq酶1U。结论Mg2+、dNTPs、引物浓度与Taq聚合酶量对不同物种SRAP-PCR结果有不同程度影响,利用SRAP分子鉴定前应对此4因素进行优化。
Objective In order to characterize Polygonum multiflorum Thunb. and its adulterants, the SRAP molecular marker system was established. Methods Orthogonal design was used to optimize the concentrations of Mg^2＋ , dNTPs, primers and Taq DNA olymerase. Results The optimum system of 20 μl was as follows ： Mg^2＋ 2.5 mmol/L, dNTPs 0.4 mmol/L, primer 0.35 μmol/L, Taq DNA polymerase 1 U. Conclusion The concentrations of Mg^2＋ , dNTPs, primers, Taq DNA olymerase have different effect in the result of SRAP - PCR in different species. It is necessary to optimize these four factors before using SRAP to identity different species.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2009年第5期1137-1139,共3页
Lishizhen Medicine and Materia Medica Research
基金
广州市科学技术基金资助项目(No.2007Z3-E5151)
