Comparison of total cycling times in amplification of genomic DNA fragments between 0.7 - 2.0kb. The increased processivity of Phusion DNA Polymerases allows extremely short cycling times. Additionally, annealing temperatures used are 5°– 8°C above conventional PCR. Both of these shave valuable seconds off of each cycle and make 2-step protocols easily achievable. The extraordinary short hold times are due to the temperature stabilizing effect of the Piko Thermal Cyclers and UTW vessels, minimizing the time required to achieve the target temperature.