Optimization and Verification of ISSR Reaction System of Eggplant
LIANG Ren-fan1, 2, WANG Yi-kui1, LI Wen-jia1, LI Yan1, WU Yong-guan1
1Institute of Vegetable, Guangxi Academy of Agricultural Sciences, Nanning 530007, Guangxi, China; 2College of Agronomy，Guangxi University, Nanning 530004 , Guangxi, China
Abstract：In this study, eggplant（Solanum melongena L.）was the test material. The total DNA was extracted from its fresh young leaves by modified CTAB method. The advantage of orthogonal design and single factor design were applied to optimize ISSR-PCR amplification system of eggplant in 4 factors, such as Mg2+, dNTP, primer and Taq DNA polymerase at 3 levels, and the optimal treatment was verified by 16 cultivate eggplants from Guangxi. The results indicated that an optimal reaction system（25 μL）was established. It was consisted of 2.5 μL 10×buffer, 2.0-3.0 mmol•L-1 Mg2+, 0.32-0.36 mmol•L-1 dNTPs, 0.4 μmol•L-1 primer, 50 ng template DNA, 0.6 U Taq DNA polymerase. The sore band of the optimized treatment showed stabilization, distinct and abundant.
