sho-phire-enzymes

Affibody-mediated hot-start PCR enzyme with improved speed, yields, and amplicons length

Thermo Scientific Phire Hot Start II DNA Polymerase is fused with a dsDNA-binding domain that allows short extension times (10–15 sec/kb) and helps improve yields compared to a standard hot-start Taq DNA polymerase. In addition, its hot-start technology with Affibody molecules allows complete activation of the enzyme in “zero-time” at standard cycling temperatures. This combination of features, along with 2x fidelity of Taq DNA polymerase, makes Phire Hot Start II DNA Polymerase an ideal solution for routine and high-throughput PCR applications.


Features:

  • Fast enzyme with quick hot start (no separate activation step required)
  • Minimal reaction optimization due to high inhibitor tolerance
  • Higher yields and amplicons length than with traditional hot-start Taq DNA polymerase
  • Direct gel loading of PCR products, with green reaction buffer formats

Complete PCR cycling in less than half the time

Phire Hot Start II DNA Polymerase provides extension time of 10 sec/kb, requires no separate activation step, and allows direct loading of PCR products onto gels. Due to these features, PCR protocols with Phire Hot Start II DNA Polymerase is significantly faster than protocols with Taq DNA polymerases.

bar chart showing short cycling times with Phire Hot Start II DNA Polymerase

 

Short cycling times with Phire Hot Start II DNA Polymerase.  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 chemical or antibody-based hot-start technologies (suppliers A-D). Green buffer further reduces experimental time by allowing fewer pipetting steps and direct gel loading.

Green reaction buffer for ultimate convenience

phire-fig2

Phire Green Hot Start II DNA Poylmerase and PCR Master Mixes are supplied with 5x Phire Green Reaction Buffer that includes a density reagent and two tracking dyes for direct loading of PCR products on gels.

Reaction mixtures containing Phire Green Reaction Buffer. (A) Before electrophoresis. (B) Blue and yellow tracking dyes after electrophoresis.  

Superior yields in significantly shorter time

Phire Hot Start II DNA Polymerase incorporates a dsDNA-binding domain that allows very short cycling times and significantly improves amplicons yields. High yields can be achieved with both Phire Hot Start II stand-alone enzyme and PCR master mix formats.

gel image showing superior target yields with Phire Hot Start II DNA Polymerase.

Superior target yields with Phire Hot Start II DNA Polymerase.  A 1.5 kb fragment from the human cathepsin K gene was amplified with 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 were substantially longer and resulted in lower product yields.

gel image showing superior target yields with Phire Hot Start II PCR Master Mix


Superior target yields with Phire Hot Start II PCR Master Mix.
A 2 kb fragment of human β-globin gene was amplified with different hot-start PCR master mixes. Phire Green Hot Start II PCR Master Mix delivered high yields of specific product in just 41 minutes whereas other master mixes provided lower yields and in most cases required longer protocols.

Higher amplicon length than with any hot start Taq DNA polymerase

Phire Hot Start II DNA Polymerase can amplify targets longer than a standard hot-start Taq DNA polymerase does. Up to 7.5 kb of gDNA and 20 kb of phage DNA can be amplified with Phire Hot Start II stand-alone enzyme and PCR master mix formats.

Phire_images_2014_Aug_4

Higher amplicons length with Phire Hot Start II DNA Polymerase. 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 high yields whereas the other hot-start Taq DNA polymerases produced significantly lower yields and failed to amplify the 7.5 kb fragment.
1: 0.6 kb, 2: 1.0 kb, 3: 1.5 kb , 4: 2.7kb, 5: 7.5 kb

Higher amplicons length with Phire Hot Start II PCR Master Mix. Five genomic DNA fragments of different lengths were amplified with hot-start PCR master mixes. Phire Green Hot Start II PCR Master Mix produced all five amplicons with high yields. Other hot-start PCR master mixes produced lower or no yields, with some also amplifying nonspecific products.
1: 0.47 kb, 2: 1.1 kb, 3: 1.7 kb, 4: 3.5 kb, 5: 7.5 kb

Complete PCR cycling in less than half the time

Phire Hot Start II DNA Polymerase provides extension time of 10 sec/kb, requires no separate activation step, and allows direct loading of PCR products onto gels. Due to these features, PCR protocols with Phire Hot Start II DNA Polymerase is significantly faster than protocols with Taq DNA polymerases.

bar chart showing short cycling times with Phire Hot Start II DNA Polymerase

 

Short cycling times with Phire Hot Start II DNA Polymerase.  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 chemical or antibody-based hot-start technologies (suppliers A-D). Green buffer further reduces experimental time by allowing fewer pipetting steps and direct gel loading.

Green reaction buffer for ultimate convenience

phire-fig2

Phire Green Hot Start II DNA Poylmerase and PCR Master Mixes are supplied with 5x Phire Green Reaction Buffer that includes a density reagent and two tracking dyes for direct loading of PCR products on gels.

Reaction mixtures containing Phire Green Reaction Buffer. (A) Before electrophoresis. (B) Blue and yellow tracking dyes after electrophoresis.  

Superior yields in significantly shorter time

Phire Hot Start II DNA Polymerase incorporates a dsDNA-binding domain that allows very short cycling times and significantly improves amplicons yields. High yields can be achieved with both Phire Hot Start II stand-alone enzyme and PCR master mix formats.

gel image showing superior target yields with Phire Hot Start II DNA Polymerase.

Superior target yields with Phire Hot Start II DNA Polymerase.  A 1.5 kb fragment from the human cathepsin K gene was amplified with 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 were substantially longer and resulted in lower product yields.

gel image showing superior target yields with Phire Hot Start II PCR Master Mix


Superior target yields with Phire Hot Start II PCR Master Mix.
A 2 kb fragment of human β-globin gene was amplified with different hot-start PCR master mixes. Phire Green Hot Start II PCR Master Mix delivered high yields of specific product in just 41 minutes whereas other master mixes provided lower yields and in most cases required longer protocols.

Higher amplicon length than with any hot start Taq DNA polymerase

Phire Hot Start II DNA Polymerase can amplify targets longer than a standard hot-start Taq DNA polymerase does. Up to 7.5 kb of gDNA and 20 kb of phage DNA can be amplified with Phire Hot Start II stand-alone enzyme and PCR master mix formats.

Phire_images_2014_Aug_4

Higher amplicons length with Phire Hot Start II DNA Polymerase. 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 high yields whereas the other hot-start Taq DNA polymerases produced significantly lower yields and failed to amplify the 7.5 kb fragment.
1: 0.6 kb, 2: 1.0 kb, 3: 1.5 kb , 4: 2.7kb, 5: 7.5 kb

Higher amplicons length with Phire Hot Start II PCR Master Mix. Five genomic DNA fragments of different lengths were amplified with hot-start PCR master mixes. Phire Green Hot Start II PCR Master Mix produced all five amplicons with high yields. Other hot-start PCR master mixes produced lower or no yields, with some also amplifying nonspecific products.
1: 0.47 kb, 2: 1.1 kb, 3: 1.7 kb, 4: 3.5 kb, 5: 7.5 kb