黄花苜蓿SSR-PCR反应体系的优化

OPTIMIZATION ON SSR-PCR REACTION SYSTEM OF MEDICAGO FALCATA L.

对黄花苜蓿SSR-PCR扩增体系的反应条件进行了优化,分析了Mg2+浓度、dNTP浓度、DNA模板浓度、引物浓度、TaqDNA聚合酶浓度对PCR扩增结果的影响。结果表明:在25μL的反应体系中,dNTPs的最佳浓度为0.2mmol/L,MgC l2的最佳浓度为2.0mmol/L,DNA模版最适加入量80ng,引物的最佳浓度为0.16μmol/L,TaqDNA聚合酶的最佳用量为1U。

In this study,we optimized SSR-PCR amplified condition on Medicago facalta L.and analyzed effects of the concentration of dNTPs,Mg^2＋,template DNA,primers and TaqDNA polymerase on PCR results.It showed that,in total volume of 25μL PCR reaction,the optimal concentration of SSR-PCR reaction system was 0.2mmol/L dNTPs,2.0mmol/L MgCl2 80ng template DNA,0.16μmol/L primers and 1U Taq DNA polymerase.