Phusion U Hot Start PCR Master Mix
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Phusion U Hot Start PCR Master Mix
Thermo Scientific™

Phusion U Hot Start PCR Master Mix

Thermo Scientific Phusion U DNA polymerase is a novel engineered high fidelity enzyme developed using fusion technology.
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Catalog NumberNo. of Reactions
F533S100 Reactions
F533L500 Reactions
Catalog number F533S
Price (USD)
320.00
Each
Add to cart
No. of Reactions:
100 Reactions
Request bulk or custom format
Price (USD)
320.00
Each
Add to cart
Thermo Scientific Phusion U Hot Start PCR Master Mix is a convenient 2X formula that contains Phusion U Hot Start DNA Polymerase, nucleotides, and an optimized reaction buffer including MgCl2.

Phusion U DNA Polymerase is a novel engineered high-fidelity enzyme developed using fusion technology. Designed with a proprietary mutation in the dUTP binding pocket of Phusion polymerase, Phusion U polymerase is able to overcome important limitations of proofreading enzymes to incorporate dUTP and read through uracil present in DNA templates.

Phusion U polymerase features properties of Phusion DNA polymerases and offers the ability to amplify long amplicons up to 20 kb and a high specificity with affibody based hot start.

These features make Phusion U Hot Start DNA Polymerase an excellent choice for such applications as amplification of bisulfite-converted or damaged DNA as well as use of carryover contamination control.

Highlights

  • High-fidelity DNA polymerase (25X Taq)
  • Engineered to incorporate dUTP and amplify uracil containing templates
  • Hot start for reduced nonspecific amplification and primer degradation
  • Only template and primers need to be added for use
  • Short extension times (15–30 s/kb)

Applications

  • Amplification of bisulfite-converted DNA
  • Amplification of damaged or aged DNA
  • Carry-over contamination control
  • Uracil-excision based (USER) cloning methods

Using Phusion DNA Polymerases

  • 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
GC-Rich PCR PerformanceMedium
PolymerasePhusion U Hot Start DNA Polymerase
Reaction SpeedFast
Product TypeHot Start PCR Master Mix
Shipping ConditionDry Ice
For Use With (Application)Hot-start PCR
Concentration2X
Fidelity (vs. Taq)25X
Hot StartBuilt-In Hot Start
No. of Reactions100 Reactions
OverhangBlunt
Reaction FormatSuperMix or Master Mix
Size (Final Product)20 kb or less
Unit SizeEach
Contents & Storage
• Phusion U Hot Start DNA Polymerase
• 2X reaction buffer
• 3 mM MgCl2
• 400 μM each of dATP, dTTP, dCTP and dGTP

Master mix is supplied with nuclease-free water.
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.