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 and Phusion Green Hot Start II High-Fidelity DNA Polymerase
Phusion and Phusion Green Hot Start II High-Fidelity PCR Master Mix
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.
20 kb fragment from λ DNA and 7.5 kb fragment from human genomic DNA was amplified with different hot start high fidelity PCR master Mixes. Phusion Green Hot Start II PCR Master Mix was the only master mix capable of providing high amounts of desired PCR products. In contrast, master mixes from other suppliers delivered zero or significantly lower yields and in some cases non-specific products.