Abstract:

In this research, annealing temperatures and several key parameters of the seedless grape SRAP-PCR reaction system were optimized. The result showed that for seedless grape, the most suitable protocol of SRAP-PCR was initially denaturing at 94℃ for 5 min, then 94℃, 1 min, 36℃ 1 min, and 72℃, 1 min for the first five cycles, and then the annealing temperature was raised to 53℃ for another 35 cycles, at last extending at 72℃ for 10 min. In this SRAP-PCR system, the optimum concentrations of Mg2+, dNTPs, Taq DNA polymerase, primers and DNA template were 1.5 mmol/L, 0.2 mmol/L, 1.0 U, 0.2 μmol/L, 40 ng/25 μL, respectively.