Establishment of the SRAPPCR reaction system and selection of the primers in Stipa bungeana

An improved CTAB method, suitable for isolating genomic DNA of Stipa bungeana, using the young leaves of this grass speicies, was established in a previous study. Based on this result, an orthogonal design and a factor analysis method were combined to optimize the SRAPPCR reaction system of S. bungeana, including five main factors: DNA, Taq Polymerase, dNTPs, Mg2+ and primers. The results indicated that the optimum concentrations of each component in the 20 L reaction system were: DNA （20 ngL-1）3 L, Taq Polymerase (5 UL-1) 0.2 L, dNTPs（2.5 mmolL-1) 1.4 L, primer（10 molL-1）1.0 L, Mg2+ (25 mmolL-1) 2.0 L, 10Buffer 2.5 L, ddH2O 8.9 L. This reaction system has been experimentally validatied and primers selected, and should be a suitable system for the genetic diversity analysis of S. bungeana. This SRAPPCR reaction system could provide a basis for the analysis of genetic diversity within S.bungeana germplasm resources, with applications being vegetation restoration and ecological reconstruction in the loess plateau.