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The optimal annealing temperature for Phusion DNA Polymerases may differ significantly from that of Taq-based polymerases.
For optimal results start by accurately calculating your Tm with our Tm calculator.
Thermo Scientific Phusion Flash PCR Master Mix was developed to save valuable laboratory time. Ready-to-use 2X master mix enables the use of extremely short PCR protocols without compromising either the fidelity or the yield in the reaction. In addition, pipetting steps are minimized: only template and primers need to be added by the user.
The unique composition of the master mix enables the use of extremely short cycling protocols with both low and high complexity DNA templates - 15 seconds per kilobase or less. The master mix utilizes Phusion Flash II DNA Polymerase, a proofreading DNA polymerase modified from Phusion Hot Start II High-Fidelity DNA Polymerase.
A 1.5 kb human cathepsin K gene was amplified with three different polymerases using varying extension times (10 to 60 s) with Thermo Scientific Piko® Thermal Cycler. Only Phusion Flash was able to amplify the 1.5 kb gene with extremely short extension times of 10 and 20 s. It also produced superior yields of specific product compared to other enzymes tested.
A 1.5 kb human cathepsin K gene was amplified with three different polymerases following manufacturer’s recommendations and using either a fast (Piko Thermal Cycler) or a conventional (DNA Engine Tetrad® from Bio-Rad) thermal cycler.