Optimization of the SRAP reaction system in broccoli and cauliflower by orthogonal design

By an orthogonal design the reaction system of brocco's SRAP(sequence-related amplified polymorphism)was optimized in terms of Mg2+,dNTPs and primers three factors and their three levels,and then the optimized SRAP reaction system was verified with 6 broccoli and cauliflower varieties.The result indicated that in a 20 μL reaction system the appropriate concentrations of dNTPs,Mg2+,and forward and reverse primers were respectively 0.2 mmolL,1.25 mmolL,0.4 μmolL and 0.4 μmolL,the DNA of template was 20 ng,and Taq DNA polymerase 1.2 U,the best annealing temperature was 50 ℃.The results offer the technique basis of using SRAP technology to study broccoli germplasm resource.