ISSR-PCR Optimization for Waxapple (Syzygium samarangense)

An optimization experiment was conducted on the waxapple cultivars, Nong Ke NO 1. Genomic DNA was extracted from the young leaves using the modified CTAB method. Orthogonal design was applied to optimize ISSR-PCR amplification of the waxapple with 5 factors, including Mg2+,dNTPs,primer,Taq DNA polymerase and template DNA, at 4 levels. An optimal reaction system (25 μL) was established, i. e., 2.0 μL 10×buffer (Mg2+ free),3.0 mmol·L-1 Mg2+,0.2 mmol·L-1 dNTPs,0.4 μmol·L-1 primer,1.5 U Taq DNA polymerase and 30 ng template DNA. Waxapple cultivars, Nong Ke NO 1 and Nong Ke NO 2, were used to challenge the optimized reaction system and select primers. The results indicated that the system was stable and 12 primers were selected.