Optimization of SSR-PCR System on Cynodon spp. by Orthongonal Design
We made an orthogonal scheme for optimizing the SSR-PCR system on Cynodon ssp.plant,which employed three levels and four factors (Mg2+,dNTP,primers,and Taq DNA polymerase,respectively) on the result of SSR and the SSR reaction was finally optimized by comparing the effects of the concentrations template DNA on application.The results showed that there are significant effects on the results of SSR-PCR under the different combinations of three levels and four factors,and the optimized SSR-PCR reaction system was generated for Cynodon spp.plant that was:10xbuffer,60 ng genomic DNA templates,25 mmol/L Mg2+,200 μmol/L dNTP,0.8 μmol/L primer,0.5 U Taq DNA polymerase with total 10 pL reaction volumes,The optimized SSR-PCR protocol might produce stable and credible banding patterns in the 10 tested Cynodon spp.populations,which could be used in genetic analysis of Cynodon spp.plant.
Cynodon spp., SSR-PCR, Orthogonal designs, System optimization
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