This calculator generates a pipetting table for setting up PCR reactions with Thermo Scientific Phusion DNA Polymerases.
The application will automatically calculate the pipetting volumes required for a premix and the volume of the premix needed for the final reaction.
*The HF buffer should be used as the default buffer for high-fidelity amplification. The GC buffer can improve the performance of Phusion DNA Polymerase on some difficult or long templates. More details can be found in the technical manuals for Phusion DNA Polymerases.
**The recommendation for final primer concentration is 0.5 µM, but it can be varied in a range of 0.2–1.0 µM, if needed.
***Addition of DMSO is recommended for GC-rich amplicons. DMSO is not recommended for amplicons with very low GC % or amplicons that are >20 kb.
For Research Use Only. Not for use in diagnostic procedures.