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 Phusion High Fidelity DNA polymerase?
Thermo Scientific Phusion High-Fidelity DNA Polymerases set a gold standard for high performance PCR. Phusion DNA Polymerases offer robust performance with short protocol times, even in the presence of PCR inhibitors, and generate higher yields with lower enzyme amounts than other DNA polymerase.
What is the difference between phusion and Taq polymerase?
Phusion is a DNA polymerase isolated from Pyrococcus furiosus and is mainly used in cloning experiments to increase the fidelity. Taq DNA Polymerase is the standard DNA polymerase used in the Polymerase Chain Reaction (PCR), and it is isolated from the thermostable bacterium, Thermus aquaticus.
Does Phusion High Fidelity DNA polymerase have proofreading?
As a proofreading enzyme with very high fidelity, Phusion Plus DNA Polymerase generates amplicons with very few errors, giving you confidence in PCR sequence accuracy.
What is Phusion buffer?
5X Thermo Scientific Phusion HF Reaction Buffer is a buffer solution optimized for high-fidelity DNA amplification using Phusion High-Fidelity DNA polymerases. The buffer contains 7.5 mM MgCl2 which provides 1.5 mM MgCl2 in final reaction conditions.
Does Phusion polymerase create blunt ends?
Phusion DNA Polymerases generate blunt end products; therefore, blunt end cloning is recommended. If TA cloning is required, it can be performed by adding A overhangs to the blunt PCR product with e.g. Taq DNA Polymerase (Cat.
What is high fidelity polymerase?
High Fidelity Polymerase is based on a blend of Taq DNA polymerase and a proofreading enzyme specially designed for highly accurate and efficient amplification. It shows excellent results with extremely long (up to 30 kb), GC-rich or other difficult templates.
Why is phusion better than Taq?
Phusion is more highly processive than Taq, so it’s faster than Taq and can handle much longer amplicons, because it has a “clamp” that keeps it attached to the DNA strand where as Taq is more likely to dissociate.
What does 5X Phusion buffer do?
5X Phusion Green Buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffers do not interfere with the enzyme performance and are compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.
Why is DMSO used in PCR?
Dimethyl sulfoxide (DMSO) is an organosulfur compound with a high polarity and high dielectric constant, that is used in PCR to disrupt secondary structure formation in the DNA template. Thus the addition of DMSO can greatly improve yields and specificities of PCR priming reactions.
Which polymerase has the highest fidelity?
Q5® High-Fidelity DNA Polymerase sets a new standard for both fidelity and robust performance. With the highest fidelity amplification available (~280 times higher than Taq), Q5 DNA Polymerase results in ultra-low error rates.
What is a low fidelity enzyme?
A superfamily of DNA polymerases that bypass lesions in DNA has been described1,2,3,4. This very low fidelity indicates a relaxed requirement for correct base pairing geometry and indicates that the function of pol-η may be tightly controlled to prevent potentially mutagenic DNA synthesis.
