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Thermo Scientific Phusion Hot Start II High Fidelity DNA Polymerase is the most accurate hot start DNA polymerase on the market. It combines the Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand. Affibody inhibits the DNA polymerase activity at room temperature and thus prevents the amplification of nonspecific products.
The Affibody ligand also inhibits the 3´→5´ exonuclease activity of the polymerase, preventing degradation of primers and template DNA during reaction set up. At polymerization temperatures, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase does not require a separate activation step in the PCR protocol as it is immediately reactivated at high temperatures.
New Phusion Green Hot Start II High-Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and 5X Phusion Green Buffers. The buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with excelent performance of Phusion Hot Start II DNA Polymerase and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.
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.
†Phusion Site-Directed Mutagenesis Kit also available.
Phusion Hot Start II High-Fidelity DNA Polymerase
Phusion Green Hot Start II High-Fidelity DNA Polymerase
Both Phusion and Phusion Green Buffers provide 1.5 mM MgCl2 in the final 1X concentration.
Five proofreading DNA polymerases from major suppliers were used to amplify 1.7 to 2.3 kb amplicons from human genomic DNA. All amplifications were performed in accordance with manufacturers instructions. Phusion Hot Start II DNA Polymerase provided high yields of specific products whereas all other enzymes delivered lower or no yields, some of them also amplifying non-specific products.