What does Phusion polymerase do?
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.
Is Pfu polymerase is thermostable?
Pfu DNA Polymerase is a high-fidelity, thermostable enzyme of approximately 90kDa isolated from Pyrococcus furiosus.
Is Phusion polymerase thermostable?
Phusion DNA Polymerase is an ideal choice for cloning and can be used for long or difficult amplicons. With an error rate > 50-fold lower than that of Taq DNA Polymerase and 6-fold lower than that of Pyrococcus furiosus DNA Polymerase (1), Phusion is one of the most accurate thermostable polymerases available.
Can PCR occur without Taq polymerase?
Taq polymerase Like DNA replication in an organism, PCR requires a DNA polymerase enzyme that makes new strands of DNA, using existing strands as templates.
What is the role of the two primers in a PCR reaction?
Two primers are used in each PCR reaction, and they are designed so that they flank the target region (region that should be copied). That is, they are given sequences that will make them bind to opposite strands of the template DNA, just at the edges of the region to be copied.
What are the guidelines for PCR using Phusion ® DNA polymerase?
The following guidelines are provided to ensure successful PCR using Phusion ® DNA Polymerase. These guidelines cover routine PCR. Amplification of templates with high GC content, high secondary structure, low template concentrations or long amplicons may require further optimization.
What is the final concentration of DNA primer in PCR?
Computer programs such as Primer3 can be used to design or analyze primers. The final concentration of each primer in a reaction using Phusion DNA Polymerase may be 0.2–1 μM, while 0.5 μM is recommended.
What is the annealing temperature of PCR polymerase?
Typically, a 5–10 second denaturation at 98°C is recommended for most templates. Annealing temperatures required for use with Phusion tend to be higher than with other PCR polymerases. The NEB Tm calculator should be used to determine the annealing temperature when using Phusion.