Thermo Scientific™ Phusion Hot Start II High-Fidelity PCR Master Mix

The most accurate hot start DNA polymerase on the market featuring extreme specificity and improved performance.

Catalog Number	Mfr. No.	No. of Reactions
F565L	Thermo Scientific™ F565L	500
F565S	Thermo Scientific™ F565S	100

Description

Phusion Hot Start II High Fidelity DNA Polymerase combines the Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand. Affibody inhibits the DNA polymerase activity at room temperature and thus prevents the amplification of nonspecific products.

The Affibody ligand also inhibits the 3‡→5‡ exonuclease activity of the polymerase, preventing degradation of primers and template DNA during reaction set up. At polymerization temperatures, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase does not require a separate activation step in the PCR protocol as it is immediately reactivated at high temperatures.

New Phusion Green Hot Start II High-Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and 5X Phusion Green Buffers. The buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with excelent performance of Phusion Hot Start II DNA Polymerase and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.

Thermo Scientific Phusion Hot Start II High Fidelity DNA Polymerase is the most accurate hot start DNA polymerase on the market. It combines the Phusion DNA Polymerase and a reversibly bound, specific Affibody ligand. Affibody inhibits the DNA polymerase activity at room temperature and thus prevents the amplification of nonspecific products.
The Affibody ligand also inhibits the 3'→5' exonuclease activity of the polymerase, preventing degradation of primers and template DNA during reaction set up. At polymerization temperatures, the ligand is released, rendering the polymerase fully active. Phusion Hot Start II DNA Polymerase does not require a separate activation step in the PCR protocol as it is immediately reactivated at high temperatures.
New Phusion Green Hot Start II High-Fidelity DNA Polymerase is a combination of Phusion Hot Start II DNA Polymerase and 5X Phusion Green Buffers. The buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with excelent performance of Phusion Hot Start II DNA Polymerase and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.


Highlights

• Reaction set up can be done at room temperature
• Reduces non-specific amplification
• Prevents primer degradation during set up
• Zero-time reactivation due to unique hot start technology
• Extreme fidelity (52X Taq)
• Speed of the polymerase allows short extension times
• Robust reactions, minimal optimization needed
• Increased product yields with minimal enzyme amounts
• Direct loading on gel with Green Buffer

Applications

• High-fidelity PCR
• High throughput
• Difficult (GC-rich) templates
• Template generation for sequencing
• Multiplex PCR
• Long-range PCR
• Cloning
• Mutagenesis†
• Microarray

†Phusion Site-Directed Mutagenesis Kit also available.

Includes
Phusion Hot Start II High-Fidelity DNA Polymerase

• Phusion Hot Start II High-Fidelity DNA Polymerase (2 U/μL)
• 5X Phusion HF Buffer
• 5X Phusion GC Buffer
• DMSO
• 50 mM MgCl₂ solution

Phusion Green Hot Start II High-Fidelity DNA Polymerase

• Phusion Hot Start II High-Fidelity DNA Polymerase (2 U/μL)
• 5X Phusion Green HF Buffer
• 5X Phusion Green GC Buffer
• DMSO
• 50 mM MgCl₂ solution

Both Phusion and Phusion Green Buffers provide 1.5 mM MgCl₂ in the final 1X concentration.

 

Hazardous: No
Shelf Life: 24 Months
Shipping Condition: Dry Ice
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 Condition: −20°C

Specifications

• Content And Storage: Includes:
  
  • 2X Phusion Hot Start II High-Fidelity PCR Master Mix (provides 1.5 mM MgCl₂ in the final 1X concentration)
  • Water, nuclease-free
  • DMSO
  
  Store at -20°C.
  
• GC-Rich PCR Performance: Medium
• Reaction Speed: Fast
• Product Type: Hot Start High-Fidelity PCR Master Mix
• Concentration: 2X
• Hot Start: Built-In Hot Start
• Reaction Format: SuperMix or Master Mix
• Starting Material: DNA

• Format: Tube
• Polymerase: Phusion Hot Start II High-Fidelity DNA Polymerase
• Product Line: Phusion
• For Use With (Application): Cloning
• Fidelity (vs. Taq): 52 X
• Overhang: Blunt
• Size (Final Product): 20 kb or Less

Current Phusion Hot Start customers can switch to Phusion Hot Start II with no changes needed to existing protocols or primers. Original Phusion Hot Start will remain available for purchase, however, all new customers are advised to purchase Phusion Hot Start II to allow more flexibility in primer design.