Abstract:

In order to find out the optimal level of four factors of melon ISSR-PCR reaction and to determine the optimum reaction system within the shortest time this study was done. The genomic DNA of melon was extracted by CTAB method and DNA sample with high purity and good integrity was obtained. In this study orthogonal design with three levels of four factors was created by software MINTAB and the analysis of variance of PCR result was carried out by MINITAB. The results showed that the primer concentration was the greatest impact on the result of the PCR reaction, Taq DNA polymerase was minimal impact on the PCR result. By the use of multiple comparison between all pairs of level a better amplification system of ISSR PCR reaction was established: 1×PCR buffer, 100 ng templates DNA, 1U Taq DNA polymerase, 0.4 μmol/L primers, 0.1mmol/L dNTP in the total volume of 20 μL.