药用菊花SRAP-PCR反应体系的优化

基于方差分析方法，利用正交试验从Taq酶、Mg2+、dNTP、引物和模板DNA等5因素4水平对药用菊花反应体系进行优化。结果标明：药用菊花SRAP-PCR的最佳反应体系为：在20μL的反应体系中，模板DNA 60ng，Taq酶1.00U，Mg2+ 2.00mmol?L-1，dNTP 0.20mmol?L-1，引物0.40μmol?L-1。Taq酶、Mg2+、dNTP和引物对SRAP反应结果有显著影响。所建立的药用菊花SRAP反应体系具有标记位点清晰、反应系统稳定、检测多态性能力较强、重复性好等特点，可用于药用菊花的遗传多样性研究。

Optimization of SRAP-PCR Reaction System for Chrysanthemum morifolium Ramat Based on the Analysis of Variance

Based on the analysis of variance,an orthogonal design was made to optimize the SRAP-PCR amplification system on Chrysanthemum morifolium Ramat using five factors(template DNA,Taq polymerase,Mg~(2+) ,dNTP and primer)at four levels of concentrations respectively.The results showed that:a suitable SRAP reaction system Was constructed with the 20 μl reaction system containing 1.00 U Taq polymerase,2.00 mmol/L Ms~(2+),0.20 mmol/L dNTP,O.40 μmol/L primer and 60ng template DNA.SRAP-PCR was significantly influenced by Taq polymerase,Ms~(2+),dNTP and primer.These SRAP-PCR systems could provide clear reliable abundant polymorphisms molecular markers and were proved suitable for the study of the genetic diversity of Chrysanthemum morifolium Ramat.