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Thermo Scientific™ Phusion™ High-Fidelity DNA Polymerase & dNTP Mix (10 mM each)

Product Code. 16237911 Shop All Thermo Scientific Products
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Product Code. 16237911 Supplier Thermo Scientific™ Supplier No. F530N

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Description

This package includes Phusion High-Fidelity DNA Polymerase (4 x 500 U) and high quality dNTP Mix (10 mM each, 2 x 1 mL).

Thermo Scientific Phusion High-Fidelity DNA Polymerase sets a high standard for high performance PCR. Featuring an error rate 52-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.

Features

  • High fidelity (52X Taq)
  • Fast PCR due to short extension times (15-30 s/kb)
  • Robust performance, minimal optimization needed
  • High yields of PCR products with minimal enzyme amounts

Applications

  • Standard PCR
  • RT-PCR
  • High fidelity and long PCR
  • LAMP, RDA, and MDA
  • cDNA synthesis
  • qPCR
  • DNA labeling and sequencing

dNTP Mix

Thermo Scientific dNTP Mix contains pre-mixed aqueous solutions of dATP, dCTP, dGTP, and dTTP, each at a final concentration of 10 mM. The nucleotides have greater than 99% purity and are free of nuclease activities, as well as human and E. coli DNA. Mixes offer the possibility of reducing the number of pipetting steps and the risk of reaction setup errors.

dNTP features

  • Stable after multiple freeze-thaw cycles
  • Up to 95% of dNTPs remain in triphosphate form after 7 weeks at room temperature
  • Up to 90% of dNTPs remain in triphosphate form after 30 cycles of PCR (1 minute at 94°C, 3 minutes at 72°C)

Notes

  • Annealing rules for Phusion DNA polymerases are different from many common DNA polymerases (such as Taq DNA polymerases).
TRUSTED_SUSTAINABILITY

Specifications

Concentration 2 U/μL
Content And Storage • Phusion High-Fidelity DNA Polymerase (4 x 500 U at 2 U/μL)
• 10 mM dNTP Mix (2 x 1 mL)
• 5X Phusion HF & GC Buffers
• DMSO
• 50 mM MgCl2 solution

Store at –5°C to –30°C.
Detection Method Primer-probe
GC-Rich PCR Performance High
PCR Method 1-step RT-qPCR
Polymerase Phusion High Fidelity DNA Polymerase
Reaction Speed Fast
Product Type High-Fidelity DNA Polymerase
Quantity 2000 reactions
Shipping Condition Dry Ice
For Use With (Application) Standard PCR, High-fidelity PCR
Fidelity (vs. Taq) 52X
Hot Start No
No. of Reactions 2000 Reactions
Overhang Blunt
Reaction Format Separate Components
Size (Final Product) 20 kb or less
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Frequently Asked Questions (FAQs)

What is enzyme concentration in Phusion Hot Start II High-Fidelity PCR Master Mix?

Phusion Hot Start II DNA polymerase concentration is optimized to give good results in most reactions. When the PCR reaction is set up according to the instructions, the final concentration of Phusion enzyme is 1 U in 50 µL reaction (0.4 U in 20 µL reaction).
Do Phusion DNA Polymerases add the non-template dependent 3'-A overhang?

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.
Can Phusion DNA Polymerases extend at 1 second/kb?

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.
Can protocols optimized for Phusion DNA Polymerase be directly applied to Phusion Hot Start II DNA Polymerase?

Yes, protocols optimized for Phusion DNA Polymerase can be applied to Phusion Hot Start II DNA Polymerase reactions.
Do Phire and Phusion Hot Start II DNA Polymerases need a separate activation step in the PCR protocol?

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.
What nucleotide analogues can I use with DyNAzyme and Phusion DNA Polymerases?

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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