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High fidelity PCR reduces sequence error rate during amplification of DNA, compared to error rate of standard PCR. High fidelity PCR uses DNA polymerases or enzyme mixes that possess 3‘→5‘ exonuclease (proofreading) activity. Such enzymes are able to correct nucleotide incorporation errors. High fidelity PCR is most important to researchers, using PCR products for cloning and expression.
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Phusion® High-Fidelity DNA Polymerase
High Fidelity PCR
Phusion® High-Fidelity DNA Polymerase
Available in certain countries only.
For availability in your country please contact your local Thermo Fisher Scientific (formerly Fermentas) office or distributor.
For availability in your country please contact your local Thermo Fisher Scientific (formerly Fermentas) office or distributor.
- Product information
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- Features
- Accuracy: The highest fidelity of any available thermostable polymerases
- Speed: Increased processivity allows shorter reaction times (extension 15-30 s/kb)
- Robustness: Fewer reaction failures and minimal optimization
- High yields: Increase product yields with minimal enzyme amounts
- Applications
- High-fidelity PCR. The error rate of Thermo Scientific Phusion DNA Polymerase is 4.4 x 10-7, which is industry-leading fidelity. Therefore, Phusion DNA Polymerase is suitable for all PCR applications requiring great accuracy.
- Cloning. Phusion DNA Polymerase amplifies templates with an accuracy and speed previously unattainable with a single enzyme. This makes it a superior choice for cloning. Phusion DNA Polymerase produces blunt ends to the amplified fragment.
- Long amplicons. Extension and overall cycling times can be significantly reduced when using Phusion DNA Polymerase. Due to the high processivity, Phusion DNA Polymerase can amplify long templates in a fraction of the time other polymerases need.
- DHPLC and Microarray applications. Detergent-free reaction buffers are available for Phusion DNA Polymerase. PCR products used for DHPLC analysis are recommended to be free of detergents. Detergents may also cause foaming when PCR products are spotted on microarray slides.
DescriptionThe unique Thermo Scientific Phusion® DNA Polymerase offers performance that no other enzyme can match. The Phusion DNA Polymerase generates templates with an accuracy and speed unattainable with a single enzyme, even on your most difficult templates. The unique structure and characteristics of Phusion DNA Polymerase make it a superior choice for cloning, and sets a new standard for PCR performance: its error rate is 50-fold lower than that of Thermus aquaticus, and 6-fold lower than that of Pyrococcus furiosus - thus being the most accurate of available thermostable polymerases. Phusion DNA Polymerase possesses processivity 10-fold greater than Pyrococcus furiosus.
Fig. 1. The schematic structure of Phusion® High-Fidelity DNA Polymerase
The dsDNA-binding domain (purple) is fused to a novel Pyrococcus-like enzyme (green) forming a unique high-performance polymerase - Phusion DNA Polymerase.
Storage Buffer20 mM Tris-HCl (pH 7.4 at 25 °C),
0.1 mM EDTA,
1 mM DTT,
100 mM KCl,
stabilizers,
200 µg/ml BSA
and 50 % glycerol.
StorageRecommended storage temperature -20 °C.
Patents, Licenses, Trademarks - Protocols & recommendations
- References
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- Frey & Suppmann, Biochemica 2: 34-35, 1995.
