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Thermo Scientific™ Phusion™ Hot Start II DNA Polymerases (2 U/μL)
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Description
Includes
Phusion Hot Start II High–Fidelity DNA Polymerase
- Phusion Hot Start II High–Fidelity DNA Polymerase (2 U/μL)
- 5X Phusion HF Buffer
- 5X Phusion GC Buffer
- DMSO
- 50 mM MgCl2 solution
Phusion Green Hot Start II High–Fidelity DNA Polymerase
- Phusion Hot Start II High–Fidelity DNA Polymerase (2 U/Glc μL)
- 5X Phusion Green HF Buffer
- 5X Phusion Green GC Buffer
- DMSO
- 50 mM MgCl2 solution
Both Phusion and Phusion Green Buffer provide 1.5 mM MgCl2 in the final 1X concentration
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.
With Phusion Hot Start II High-Fidelity DNA Polymerase 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' to 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 Hot Start II High-Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and the 5X Phusion buffers. The buffers include a density reagent and two tracking dyes which do not interfere with the robust performance of Phusion Hot Start II DNA Polymerase. Researchers may directly load PCR products on a gel making this a versatile option for use in downstream applications such as DNA sequencing, ligation and restriction digestion.
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
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).
Specifications
Specifications
| Concentration | 2 U/μL |
| Content And Storage | • Phusion Hot Start II High-Fidelity DNA Polymerase (2 U/μL) • 5X Phusion Green HF Buffer • 5X Phusion Green GC Buffer • DMSO • 50 mM MgCl2 solution Phusion Green HF and GC Buffers both provide 1.5 mM MgCl2 in the final 1X concentration. Store at –20°C. |
| GC-Rich PCR Performance | High |
| Polymerase | Phusion Hot Start II High-Fidelity DNA Polymerase |
| Reaction Speed | Fast |
| Product Type | Hot Start DNA Polymerase |
| Quantity | 100 units |
| Shipping Condition | Dry Ice |
| For Use With (Application) | Hot-start PCR, High-fidelity PCR |
| Fidelity (vs. Taq) | 52X |
| Show More |
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.
No separate activation step is required since Phire and Phusion Hot Start II DNA Polymerases are inactivated by a reversibly bound, specific Affibody ligand that dissociates during initial denaturation.
DyNAzyme II DNA Polymerase can use dUTP, biotinylated dNTPs, 7-deaza-dGTP, digoxigenin-dUTP, bromo-dUTP, radiolabeled dNTPs and ITP. DyNAzyme EXT DNA Polymerase and Phusion DNA Polymerase cannot read dUTP-derivatives or dITP in the template strand so the use of these analogues is not recommended. Use Phusion U Hot Start DNA Polymerase for amplification of dUTP and dITP containing templates.
For Research Use Only. Not for use in diagnostic procedures.
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