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 is Phusion polymerase used for?
Phusion polymerase is used to obtain extreme fidelity over the conventional thermostable polymerase; Taq polymerase. Phusion polymerase is capable of amplifying long templates up to 7.5kb of genomic DNA. The optimal polymerization capacity of Phusion polymerase is at 720 C.
How do you dilute Phusion polymerase?
For example, to increase the MgCl2 concentration to 2.0 mM, add 0.5 µL of the 50 mM MgCl2 solution. Because the PCR reactions can be quite sensitive to changes in the MgCl2 concentration, it is recommended that the 50 mM MgCl2 stock solution is diluted 1:5 (to 10 mM) to minimize pipetting errors.
What is Phusion High-Fidelity DNA Polymerase?
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.
Is Phusion a proofreading polymerase?
Phusion Polymerase: robust and accurate high-fidelity PCR The enzyme, originally developed by Finnzymes, is a fusion of a proofreading polymerase and a processivity-enhancing domain, creating one of the most reliable and robust polymerases available.
Is Phusion High Fidelity?
For high speed and high performance PCR. Manufactured and quality-controlled at New England Biolabs, Thermo Scientific® Phusion High-Fidelity DNA Polymerase offers both high fidelity and robust performance.
What is the annealing temperature?
The annealing temperature of a standard PCR protocol is either 55°C [2, 3] or 60°C [4]. The chosen temperature depends on the strand-melting temperature of the primers and the desired specificity.
What does phusion mean?
Phusion is a high-fidelity DNA polymerase that offers extreme performance for all PCR applications. Incorporating an exciting new technology developed in collaboration with MJ Bioworks, Inc., Phusion DNA polymerase brings together a novel Pyrococcus-like enzyme with a processivity- enhancing domain.
How much DMSO do I add to PCR?
PCR additives: The recommended reaction condition for GC-rich templates include 3% DMSO as a PCR additive, which aids in the denaturing of templates with high GC contents.
What does DMSO do to DNA?
DMSO binds to the DNA and prevents the reannealing of single-stranded DNA. It also facilitates the annealing of primer with a templet. Therefore, it increases the specificity and yield of the PCR reaction. Generally, the GC content of the template DNA for PCR is between 45% to 52%.