Description:
High Fidelity DNA Polymerases are important for applications in which the DNA sequence needs to be correct after amplification. Manufactured and quality controlled at New England Biolabs, Thermo Scientific® Phusion High- Fidelity DNA Polymerase offers both high fidelity and robust performance, and thus can be used for all PCR applications. Its unique structure, a novel Pyrococcus-like enzyme fused with a processivityenhancing 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 High-Fidelity PCR Master Mix with GC Buffer offers robust performance and can be used for all PCR applications. It is a 2X master mix consisting of Phusion DNA Polymerase, deoxynucleotides and reaction buffer that has been optimized and includes MgCl2. All that is required is the addition of template, primers and water.
Source:
An E. coli strain that carries the Phusion DNA Polymerase gene. Applications:
PCR
Cloning
Long or Difficult Amplification
High-Throughput PCR
Reagents Supplied:
DMSO (100 %)
Heat Inactivation:
No
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C. Unit Assay Conditions:
25 mM TAPS-HCl (pH 9.3 @ 25°C), 50 mM KCl, 2 mM MgCl2, 1 mM β-mercaptoethanol, 200 µM dNTPs including [3H]-dTTP and 400 µg/ml activated Calf Thymus DNA. Reaction Conditions:
1X Phusion High-Fidelity PCR Master Mix with GC Buffer, DNA template, 0.5 µM primers and 3% DMSO (optional) in a total reaction volume of 50 µl. Phusion 1X Master Mix:
20 units/ml Phusion DNA polymerase
0.2 mM each dNTP
1X Phusion GC Buffer