Abstract:

To established a SRAP-PCR reaction system for the grape 5BB. In this paper, the orthogonal design was used to optimize SRAP-PCR amplification system on grape on four levels of five factors (Taq DNA polymerase, Mg2+, DNA template, dNTPs, and primer, respectively). The results showed the order of each factor in different levels affected on the result of PCR was Mg2 + , primer, dNTPs,Taq DNA polymerase and DNA template. A most suitable SRAP-PCR system for grape 5BB was established that was total 20 μL reaction system containing 2 UTaq DNA polymerase, 2.0 mmol/L Mg2+, 60 ng DNA template, 0.25 mmol/L dNTPs, and 0.10 μmol/L primer. This optimized system for SRAP marker would become one of the protocols for further research on grape.