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Description
Thermo Scientific Phusion High-Fidelity DNA Polymerase sets a high 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 an 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 polymerases.
The Phusion High-Fidelity PCR Kit contains all the necessary reagents for PCR including control lambda DNA template and primers for 1.3 kb and 10 kb amplicons. The DNA template amount is sufficient for performing 20 control reactions in 50 μL volume or 50 control reactions in 20 μL volume.
Highlights
- High fidelity (52X Taq)
- Fast PCR due to short extension times (15–30 s/kb)
- Robust performance, minimal optimization needed
- Increased product yields with minimal enzyme amounts
Applications
- High-fidelity PCR
- High throughput
- Amplification of difficult (GC-rich) templates
- Template generation for sequencing
- Multiplex PCR
- Long-range PCR
- Cloning & mutagenesis
- Microarray
Notes
- Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases (such as Taq DNA polymerases).
Specifications
Specifications
| Content And Storage | • Phusion DNA Polymerase (2 U/μL) • 5X Phusion GC & HF Buffer • dNTP mix (10 mM each) • Control template (λ DNA) and DNA size standard • 1.3 kb & 10 kb primers (4 μM each) • 50 mM MgCl2 solution and DMSO Store at –20°C. |
| GC-Rich PCR Performance | High |
| Polymerase | Phusion High-Fidelity DNA Polymerase |
| Reaction Speed | Fast |
| Product Type | High-Fidelity PCR Kit |
| Quantity | 50 reactions |
| Shipping Condition | Dry Ice |
| For Use With (Application) | Standard PCR, High-fidelity PCR |
| Fidelity (vs. Taq) | 52X |
| Hot Start | No |
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Frequently Asked Questions (FAQs)
Phusion DNA Polymerases generate blunt end products; therefore, blunt end cloning is recommended. If TA cloning is required, it can be performed by adding A overhangs to the blunt PCR product with e.g. Taq DNA Polymerase (Cat. No. EP0401). However, before adding the overhangs it is very important to remove all the Phusion DNA Polymerase by purifying the PCR product carefully, as the proofreading activity in Phusion DNA Polymerase is very strong. Any remaining Phusion DNA Polymerase will degrade the A overhangs, thus creating blunt ends again.
Yes it is possible, especially when amplifying smaller amplicons. Processivity of Phusion DNA Polymerases is 10 times that of Pfu. We recommend extension times of 15 s/kb for Phusion Flash PCR Master Mix. 15 s/kb is a conservative value that we can promise to work with almost any amplicon. In many cases, significantly shorter extension times (0-5 s/kb) can be used without compromising the yield. What separates Phusion Flash DNA Polymerase from other fast polymerases is that all steps in the PCR protocol can be shortened, including annealing and denaturation. This results in extremely fast protocols as compared with any other polymerase.
Yes, protocols optimized for Phusion DNA Polymerase can be applied to Phusion Hot Start II DNA Polymerase reactions.
For Research Use Only. Not for use in diagnostic procedures.
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