Optimization of ISSR-PCR system on Zoysia spp.

The ISSR-PCR system was optimized and the effects of template DNA with different concentrations on the PCR amplification were studied by an orthogonal experimental design with 3 different levels of 4 factors,e.g.Mg2+,dNTP,primers,and Taq DNA polymerase.The results show that each factor in different levels had significant effect on the amplification result of PCR and the orthogonal design was useful to optimize the ISSR-PCR system.The optimized PCR reaction system for Zoysia spp.was 10X Buffer,70 ng genomic DNA template,2.0 mmol·L-1 Mg2+,150 μmol·L-1 dNTP,0.3 μmol·L-1 Primer,1.0 U Taq DNA polymerase with total reaction solution of 20 μL.The system was proved stable and credible according to the result of 11 Zoysia spp.and the result of this study could be useful to provide the basis in the analysis of genetic diversity,germplasm identification,and kinship of Zoysia spp.