2023 Volume 49 Issue 2 Pages 41-48
DNA from a total of 7 mushroom species (5 orders, 5 families) was extracted and PCR amplified using existing primers of atp6. Although these taxa had a previous record of highly successful PCR amplifications of atp6, the results from an initial attempt ranged from a complete failure to a moderate success. When using slightly modified PCR cycle parameters, especially with a reduced ramp rate for increasing the temperature from the annealing (37°C) to the extension (72°C), a success rate of PCR became significantly higher in most cases. However, the results varied depending on the type of thermal cyclers used. In an extreme case, the thermal cycler by one manufacturer did not produce even a single positive band. Our study strongly indicates that a simple adjustment of the ramp options could make a significant difference in PCR amplifications without changing other important factors, such as polymerases and primer designs.
