How does Phusion polymerase work?

How does Phusion polymerase work?

Phusion DNA Polymerase possesses 5´→ 3´ polymerase activity, 3´→ 5´ exonuclease activity and will generate blunt-ended products. A 3.8 kb fragment was amplified from 50 ng of Jurkat gDNA using different polymerases. Reactions were carried out according to the manufacturer’s recommended conditions.

What is phusion mix?

Phusion High-Fidelity PCR Master Mix with HF Buffer is a 2X master mix consisting of Phusion DNA Polymerase, deoxynucleotides and reaction buffer that has been optimized and includes MgCl2. All that is required is the addition of template, primers and water.

What is Phusion GC buffer?

Create optimal reaction conditions for high fidelity amplification. The default buffer for use with Phusion® High Fidelity DNA Polymerase. Supplied at 5X concentration.

Where did Taq polymerase come from?

function in polymerase chain reaction …a heat-stable DNA polymerase called Taq, an enzyme isolated from the thermophilic bacterium Thermus aquaticus, which inhabits hot springs. Taq polymerase also led to the invention of the PCR machine.

What is Phusion PCR?

Phusion DNA Polymerase brings together a novel Pyrococcus-like enzyme with a processivity-enhancing domain and generates PCR products with accuracy and speed previously unattainable with a single enzyme, even on your most difficult templates. Additionally, Phusion DNA Polymerase is capable of amplifying long templates.

What type of polymerase is phusion?

DNA Polymerase
Thermo Scientific Phusion High-Fidelity DNA Polymerases set a gold standard for high performance PCR.

Who discovered Thermus aquaticus?

Dr. Thomas Brock
In 1967, microbiologist Dr. Thomas Brock and undergraduate student Hudson Freeze (Indiana University) were able to isolate a novel bacteria, later named Thermus aquaticus, in the Lower Geyser Basin of YNP.

Is Phusion High Fidelity?

Thermo Scientific Phusion High-Fidelity DNA Polymerases set a gold standard for high performance PCR. Featuring an error rate 50-fold lower than that of Taq, and 6-fold lower than that of Pfu, Phusion High-Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity.

Which buffer is used in the Phusion high-fidelity PCR master mix?

This Master Mix contains Phusion HF Buffer. HF buffer is recommended as the default buffer for high-fidelity amplification. For difficult templates, such as GC-rich templates or those with secondary structure, Phusion High-Fidelity PCR Master Mix with GC Buffer ( NEB #M0532) can improve reaction performance.

What is the difference between HF buffer and GC buffer?

HF buffer is recommended as the default buffer for high-fidelity amplification. For difficult templates, such as GC-rich templates or those with secondary structure, Phusion High-Fidelity PCR Master Mix with GC Buffer ( NEB #M0532) can improve reaction performance. GC buffer should be used in experiments where HF buffer does not work.

What is the final concentration of Phusion DNA polymerase?

If reactions are set up according to recommendations listed, the final concentration of Phusion DNA Polymerase in the reaction is 1 unit/50 µl or 0.4 units/20 µl. This Master Mix contains Phusion HF Buffer. HF buffer is recommended as the default buffer for high-fidelity amplification.

Can Phusion master mix be used with dUTP?

Mg ++, deoxynucleotides and additives: At 1X concentration, Phusion Master Mix provides 1. 5 mM MgCl 2 and 200 µM of each dNTP in the final reaction. Phusion cannot incorporate dUTP and is not recommended for use with uracil-containing primers or template.