Phusion High-Fidelity PCR Kit
Phusion High-Fidelity PCR Kit
Thermo Scientific™

Phusion High-Fidelity PCR Kit

Thermo Scientific Phusion High-Fidelity DNA polymerases set a gold standard for high performance PCR.
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Catalog NumberNo. of Reactions
F553L200 Reactions
F553S50 Reactions
Catalog number F553L
Price (USD)
294.65
Online Exclusive
353.00
Save 58.35 (17%)
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No. of Reactions:
200 Reactions
Request bulk or custom format
Price (USD)
294.65
Online Exclusive
353.00
Save 58.35 (17%)
Each
Add to cart
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).
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Fidelity (vs. Taq)52X
Hot StartNo
No. of Reactions200 Reactions
OverhangBlunt
PolymerasePhusion High-Fidelity DNA Polymerase
Product TypeHigh-Fidelity PCR Kit
Quantity200 reactions
Reaction FormatSeparate Components
Shipping ConditionDry Ice
Size (Final Product)20 kb or less
For Use With (Application)Standard PCR, High-fidelity PCR
GC-Rich PCR PerformanceHigh
Reaction SpeedFast
Unit SizeEach
Contents & 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.

Frequently asked questions (FAQs)

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.