What is in Phusion master 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.
Why is Phusion such a powerful PCR Polymerase?
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. Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases (such as Taq DNA polymerases).
What is the rate at which the Phusion Flash II DNA Polymerase extends DNA?
15 s/kb
Processivity of Phusion DNA Polymerases is 10 times that of Pfu. We recommend extension times of 15 s/kb for Phusion Flash PCR Master Mix. 15 s/kb is a conservative value that we can promise to work with almost any amplicon.
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.
What is phusion mix?
How much DMSO do you add to PCR?
Generally, the GC content of the template DNA for PCR is between 45% to 52%. If the GC content is higher than the desired range use 5% DMSO in PCR reaction. 4% to 10% DMSO concentration can be utilized to optimize the PCR reaction.
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 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 GC buffer?
4x GC Buffer I and 4x GC Buffer II is developed to promote excellent amplification in combination with TEMPase Hot Start DNA Polymerase of targets with varying degrees of GC content. Promotes amplification of GC-rich DNA. 4x PCR buffer. Promotes diminished formation of non-specific PCR products.
What does magnesium do in PCR?
Magnesium ion (Mg2+) functions as a cofactor for activity of DNA polymerases by enabling incorporation of dNTPs during polymerization. The magnesium ions at the enzyme’s active site catalyze phosphodiester bond formation between the 3′-OH of a primer and the phosphate group of a dNTP (Figure 6).
What is Phusion high fidelity PCR master mix?
Phusion High-Fidelity PCR Master Mix is convenient 2X mix containing Phusion DNA Polymerase, nucleotides and optimized reaction buffer including MgCl 2. Two master mix formulations are available – with HF Buffer (F-531S and F-531L) and with GC Buffer (F-532S and F-532L).
Why choose a Phusion DNA polymerase?
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. Phusion High-Fidelity PCR Master Mix is convenient 2X mix containing Phusion DNA Polymerase, nucleotides and optimized reaction buffer including MgCl 2.
What is Thermo Scientific Phusion high-fidelity DNA polymerase?
Manufactured and quality-controlled at New England Biolabs, Thermo Scientific ® Phusion High-Fidelity DNA Polymerase offers both high fidelity and robust performance, and thus can be used for all PCR applications. Its unique structure, a novel Pyrococcus -like enzyme fused with a processivity-enhancing domain, increases fidelity and speed.
What is the pH of Phusion hot start flex DNA polymerase?
25 mM TAPS-HCl (pH 9.3 @ 25°C), 50 mM KCl, 2 mM MgCl 2 , 1 mM β-mercaptoethanol, 200 µM dNTPs including [ 3 H]-dTTP and 15 nM primed M13 DNA. Phusion Hot Start Flex DNA Polymerase is unlike other enzymes and care must be taken when designing cycling protocols.