Optimization of ISSR reaetion system in pitaya

In order to establish a stable ISSR-PCR reaction system for pitaya,a single factor experiment and orthogonal method was applied for the optimization.The results showed that the optimum concentrations of different components in 20 μL reaction system were 2.0 μL 10×Buffer(with Mg2+),20.0 ng template DNA,0.30 mmolL-1 dNTP,0.35 μmolL-1 primer and 1.5 U Taq DNA polymerase.The optimized annealing temperature was 54.0℃ for the primer(AC)8T.The established ISSR-PCR system was tested on 6 pitaya samples with satisfactory performance.