Phire Hot Start II DNA Polymerase


Phire Hot Start II DNA Polymerase
A novel hot start DNA Polymerase for routine and high throughput PCR applications requiring speed and specifity.
  
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Phire Hot Start PCR II DNA Polymerase StructureThermo Scientific Phire Hot Start II DNA Polymerase is a novel PCR enzyme for routine and high throughput PCR applications. It outperforms every Taq-based hot start polymerase on the market. Phire Hot Start II DNA Polymerase is significantly faster, extremely robust, and also capable of amplifying long DNA fragments with high yields. These features are achieved through advanced protein engineering of the polymerase. It incorporates a unique double-stranded DNA binding domain which allows short extension times (10 to 15 s/kb), improves yields, and increases fidelity 2-fold compared to Taq DNA polymerase.

The hot start modification of Phire Hot Start II DNA Polymerase is based on the same Affibody inactivation technology utilized by Phusion Hot Start II DNA Polymerase. This technology increases the specificity of the PCR reaction with no additional time required for initial activation of the enzyme.

New Phire Green format is a combination of Phire Hot Start II DNA Polymerase and 5X Phire Green Buffer. The buffer includes a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffer does not interfere with enzyme performance and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.

Phire PolymerasesDID YOU KNOW?

The optimal annealing temperature for Phire DNA Polymerases may differ significantly from that of Taq-based polymerases.

For optimal results start by accurately calculating your Tm with our Tm calculator.

Highlights

  • Quick hot start – No reactivation step
  • Fast enzyme – Amplify four times faster than with hot start Taq
  • Robust – Minimal reaction optimization due to high inhibitor tolerance
  • High yields – Abundant products due to high efficiency
  • Longer PCR products – Amplify significantly longer DNA fragments than with any hot start Taq
  • Direct loading on gel with Green Buffer

Applications

  • Hot Start PCR
  • Routine PCR
  • Non-high fidelity PCR
  • Fast PCR
  • High throughput PCR
  • Genotyping

Includes

Phire Hot Start II DNA Polymerase

  • Phire Hot Start II DNA Polymerase
  • 5X Phire Reaction Buffer (provides 1.5 mM MgCl2 in the final 1X concentration)
  • DMSO

Phire Green Hot Start II DNA Polymerase

  • Phire Hot Start II DNA Polymerase
  • 5X Phire Green Reaction Buffer (provides 1.5 mM MgCl2 in the final 1X concentration)
  • DMSO
  
HazardousNo
Shelf Life24 Months
Shipping ConditionDry Ice
Storage Condition-20 C
Competitor comparison

Superior yields in significantly shorter time

Competitor comparison

A 1.5 kb fragment from the human cathepsin K gene was amplified with five different hot start DNA polymerases. Phire Hot Start II DNA Polymerase amplified high amounts of specific PCR product in just 29 minutes. In contrast, the PCR protocols for hot start Taq DNA polymerases from four major suppliers (A through D) were substantially longer and resulted in lower product yields.


Competitor comparison

Phire Hot Start II amplifies longer fragments than any hot start Taq

Competitor comparison

Five genomic DNA fragments of different lengths were amplified with three different hot start DNA polymerases. Phire Hot Start II DNA Polymerase produced all five amplicons with very high yields. The competing hot start Taq DNA polymerases produced significantly lower yields and failed to amplify the 7.5 kb fragment.

1 – 0.6 kb fragment of human glutathione peroxidase 3 gene
2 – 1.0 kb fragment of human glutathione peroxidase 3 gene
3 – 1.5 kb fragment of human cathepsin K gene
4 – 2.7 kb fragment of human Β-2-microglobulin gene 5 to 7.5 kb fragment of human Β-globin gene


Comparison of PCR cycling times

Complete PCR protocols in less than half the time.

Comparison of PCR cycling times

A 600 bp fragment from human genomic DNA was amplified with five different hot start DNA polymerases. With Phire Hot Start II DNA Polymerase, the PCR protocol was completed up to four times faster than with Taq DNA polymerases utilizing chemically modified or antibody-based hot start technologies (suppliers A through D).