Optimization of ISSR - PCR Reaction System of Fontinaloideae

Objective: The ISSR-PCR reaction system was optimized to lay the foundation for genetic diversity of Fontinaloideae.Method: Genomic DNA was extracted from Fontinaloideae,five factors and four levels of ISSR-PCR reaction system were optimized with the orthogonal experimental method,and the impact of the five factors for PCR amplification was observed based on calculating variance analysis.Result: The best ISSR-PCR reaction system of Fontinaloideae contained 2.0μL PCR buffer,1.52mmol.L-1 Mg 2 +,1.28μmol.L-1 primer,1.20mmol.L-1 dNTPs,70.0 ng polymerase,0.50 U Taq polymerase.The effect order of five factors was Mg 2 + ﹥ Taq poly-merase ﹥ dNTPs ﹥ template DNA = primer.Conclusion: The best reaction system was obtained by isolated genomic DNA and optimized the system,which had laid the good foundation for ISSR analysis on studies of Fontinaloideae resources identification and genetic diversity.