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Phusion® Hot Start II

Phusion® Hot Start II High-Fidelity DNA Polymerase


  Phusion® Hot Start II
 


Improved version of the most accurate hot start DNA polymerase available

Proofreading DNA polymerase with superior features

Finnzymes’ Phusion® Hot Start High-Fidelity DNA Polymerase is the most accurate hot start DNA polymerase in the market. It has now been upgraded to allow the use of even wider range of primers, including those with low melting temperatures. The new Phusion® Hot Start II DNA Polymerase is ideal for PCR applications from routine PCR to highly demanding applications such as cloning and high-throughput PCR. Like all Phusion DNA Polymerases, it incorporates a unique dsDNA-binding domain making this Pyrococcus-like proofreading DNA polymerase extremely robust, rapid, and accurate.

Advantages

  • No annealing temperature limitations
  • Allows reaction setup at room temperature
  • Zero-time reactivation due to unique hot start technology
  • Fast PCR enzyme with extreme fidelity (over 6 times more accurate than Pfu)
  • High product yields with low enzyme amounts gives excellent cost/reaction ratio
  • Robust enzyme with high tolerance against PCR inhibitors requires minimal optimization

Extreme specificity with improved performance

At room temperature Phusion Hot Start II DNA Polymerase is inactivated by a reversibly binding Affibody® molecule, which makes the enzyme extremely specific. Hot start modification also prevents primer degradation during reaction setup. The unique hot start technology allows zero-time reactivation of the enzyme at standard cycling temperatures. The new Affibody molecule utilized in the Phusion Hot Start II DNA Polymerase sets no temperature limitations in the annealing step of the PCR protocol. This improvement allows the use of a very wide range of primers, including those with low melting temperatures.

Gel image comparing Phusion Hot Start II DNA Polymerase to other suppliers' polymerases

Phusion Hot Start II DNA Polymerase provides extreme specificity and abundant yields. Five proofreading DNA polymerases from major suppliers were used to amplify 1.7-2.3 kb amplicons from human genomic DNA. All amplifications were performed in accordance with manufacturers’ instructions. Phusion Hot Start II DNA Polymerase provided high yields of specific products whereas all other enzymes delivered lower or no yields, some of them also amplifying non-specific products.

General information about Phusion DNA Polymerases, their advantages and applications can be found on Phusion DNA Polymerase products page.

Contents

Phusion Hot Start II High-Fidelity DNA Polymerase is supplied with 5x Phusion HF Buffer, 5x Phusion GC Buffer, DMSO and 50 mM MgCl2 solution. Both Phusion HF Buffer and Phusion GC Buffer contain 7.5 mM MgCl2 in the provided 5x concentration.

Storage buffer
20 mM Tris-HCl (pH 7.4 at 25°C), 0.1 mM EDTA, 1 mM DTT, 100 mM KCl, stabilizers, 200 µg/ml BSA and 50% glycerol.

Storage and stability

Recommended storage temperature -20°C. Stable for one year from the assay date.

Ordering information

NEW Phusion Hot Start II DNA Polymerase:
Phusion® Hot Start II High-Fidelity DNA Polymerase

F-549S

100 U (2 U/µl)

F-549L

500 U (2 U/µl)

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