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  7. Thermo Scientific™ Phusion Flash High-Fidelity PCR Master Mix

Thermo Scientific™ Phusion Flash High-Fidelity PCR Master Mix

Catalog No. F548S
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F548L 500 Reactions
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Description

Thermo Scientific Phusion Flash PCR Master Mix was developed to save valuable laboratory time.

Thermo Scientific Phusion Flash PCR Master Mix was developed to save valuable laboratory time. Ready-to-use 2X master mix preserves the fidelity and the yield in reactions when using very short PCR protocols. The unique master mix composition permits usage of short cycling protocols with both low and high complexity DNA templates (15 seconds per kilobase or less). The master mix utilizes Phusion Flash II DNA Polymerase, a modified proofreading DNA polymerase derived from Phusion Hot Start II High-Fidelity DNA Polymerase.

Features

  • Extreme speed with extension times of 15 s/kb or less
  • Hot start modification allows 'zero-time reactivation'
  • Accurate proofreading DNA polymerase with a fidelity of 25X Taq
  • High yields in reduced cycling time
  • Minimized pipetting steps

Applications

  • Fast PCR
  • High-fidelity PCR
  • 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

Concentration 2X
Content And Storage Phusion Flash High-Fidelity PCR Master Mix (1 mL)

Store at -20°C.
GC-Rich PCR Performance High
Polymerase Phusion Flash II DNA Polymerase
Reaction Speed Fast
Product Type High-Fidelity PCR Master Mix
Quantity 100 reactions
Shipping Condition Dry Ice
For Use With (Application) Hot-start PCR, High-fidelity PCR
Fidelity (vs. Taq) 25X
Hot Start Built-In Hot Start
No. of Reactions 100 Reactions
Overhang Blunt
Reaction Format SuperMix or Master Mix
Size (Final Product) 20 kb or less
Color Colorless
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Frequently Asked Questions (FAQs)

What is the difference between Phusion Flash High-Fidelity PCR Master Mix and Phusion Hot Start II High-Fidelity PCR Master Mix?

Phusion Hot Start II High-Fidelity PCR Master Mix is a 2X ready-to-use solution based on Phusion Hot Start II DNA Polymerase. It is designed for the highest fidelity (52X Taq) and specificity. Phusion Flash High-Fidelity PCR Master Mix is based on a modified Phusion Hot Start II DNA Polymerase, which allows for extremely short cycling times and features somewhat lower fidelity (25X Taq).
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.

For Research Use Only. Not for use in diagnostic procedures.