Thermo Scientific Phusion High-Fidelity DNA Polymerases set a gold standard for high performance PCR. Featuring an error rate 50-fold lower than that of Taq, and 6-fold lower than that of Pfu, Phusion High-Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity. Phusion DNA Polymerases offer robust performance with short protocol times, even in the presence of PCR inhibitors, and generate higher yields with lower enzyme amounts than other DNA polymerase.
Highlights
Reaction set up at room temperature
No non-specific amplification and primer degradation during reaction set up
Zero-time reactivation due to unique hot start technology
High fidelity (52X Taq)
Fast PCR due to short extension times (15-30 s/kb)
Robust reactions, minimal optimization needed
Increased product yields with minimal enzyme amounts
Direct loading on gels
With Phusion Hot Start II High-Fidelity DNA Polymerases amplification proceeds without the production of nonspecific products due to the combination of Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand that inhibits DNA polymerase activity at room temperature. The Affibody ligand also inhibits the 3'→5' exonuclease activity of the polymerase, thus preventing degradation of primers and template DNA during reaction set up. At temperatures that promote polymerase activity, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase is immediately reactivated at high temperatures so it does not require a separate activation step in PCR protocols.
Phusion Green Hot Start II High-Fidelity PCR Master Mix is convenient 2X mix designed to minimize the number of pipetting steps. The master mix contains Phusion Hot Start II DNA Polymerase, nucleotides and optimized reaction buffer including MgCl2. The buffer also includes a density reagent and two tracking dyes for direct loading of PCR products on a gel.
Applications
High-fidelity PCR
High throughput
Amplification of difficult (GC-rich) templates
Template generation for sequencing
Multiplex PCR
Long-range PCR
Cloning
Mutagenesis
Microarray
Using Phusion DNA Polymerases
Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases (such as Taq DNA polymerases).
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