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Reaction setup with Phusion® DNA Polymerases
This calculator generates a pipetting table for setting up PCR reactions with Thermo Scientific Phusion® DNA Polymerases.
- Fill in the volume of template DNA you want to use, the number of PCR reactions you intend to run, and the individual reaction volume.
- Define if you would like to add some extra volume for pipetting. You can choose between an absolute volume, a number of extra reactions or a presentage of the final volume.
- Fill in your stock concentrations and the desired final concentrations for primers, dNTPs, (DMSO) and Phusion DNA Polymerase.
The application will automatically calculate the pipetting volumes required for a premix, and the volume of the premix needed for the final reaction.
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| Reagents | Instruments | Consumables |
| * | 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. See more details in the Phusion instruction manuals. |
| ** | 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. |