真菌のSSR-PCR条件の適正化に関する研究

抄録

Objective: To investigate the influential factors of SSR-PCR experiments, the optimal reaction system in fungi was established, which was based for the further research on fungal identification and genetic diversity. Methods: The orthogonal design was used to optimize SSR-PCR amplification system of fungi with four factors (Taq DNA polymerase, DNA template, dNTP and primer). The optimal annealing temperature and cycles of SSR-PCR for fungal reaction were proposed. Results: The notable influence of each factor in different levels was found and the quantity of DNA template was the most effective factor. It is 25μl reaction system which contained 1.0U Taq DNA polymerase, 30ng DNA template, 150μmol/L dNTP, 0.25 μmol /L primer. Conclusion: The method of orthogonal design had been invented to solve multi-factor-controlling problems. It is simple, rapid and efficient to get the optimal conditions of PCR which is an effective method deserving to spread.