High-Fidelity DNA Polymerases are important for applications in which the DNA sequence needs to be correct after amplification. Its unique structure, a novel Pyrococcus-like enzyme fused with a processivity-enhancing domain, increases fidelity and speed. Phusion DNA Polymerase is an ideal choice for cloning and can be used for long or difficult amplicons. With an error rate > 50-fold lower than that of Taq DNA Polymerase and 6-fold lower than that of Pyrococcus furiosus DNA Polymerase (1), Phusion is one of the most accurate thermostable polymerases available. Phusion DNA Polymerase possesses 5´→ 3´ polymerase activity, 3´→ 5´ exonuclease activity and will generate blunt-ended products.
Phusion DNA Polymerase is supplied with standard 5X Phusion HF Buffer, as well as 5X Phusion GC Buffer, which can be used for complex or GC-rich templates. Each of these buffers contains MgCl2 (1.5 mM at the final [1X] reaction concentration). Reactions can also be optimized using the provided DMSO or MgCl2 solutions.
Fidelity assays were performed using a lacI-based method modified from Frey & Suppman, 1995.A 3.8 kb fragment was amplified from 50 ng of Jurkat gDNA using different polymerases. Reactions were carried out according to the manufacturer"s recommended conditions. Extension times are indicated (in minutes). Ladder L is a 1 kb DNA Ladder (NEB# N3232).
Product Source
AnE. colistrain that carries the Phusion DNA Polymerase gene.
This product is related to the following categories:
Phusion® High-Fidelity DNA Polymerases Products
This product can be used in the following applications: