Phusion dna polymerase may be diluted in 1x hf or gc buffer just prior to use in order to reduce pipetting errors. Dna polymerases with high fidelity are important for applications in which the dna sequence needs to be correct after amplification. L total if you are using any of the phusion pcr master mix products, add 25 or 10. Phusion hot start ii highfidelity dna polymerase is capable of amplifying long amplicons such as the 7. Phusion dna polymerase is an ideal choice for cloning and can be used for long or difficult amplicons. The thermo scientific phusion highfidelity dna polymerase offers high performance for all major pcr applications. Highfidelity dna polymerase offers both high fidelity and robust performance, and thus can be used for all pcr. Fret image correlation spectroscopy reveals rnapii. The phusion highfidelity pcr kit contains a sufficient supply of phusion highfidelity dna polymerase, phusion hf and gc buffers, deoxynucleotides, magnesium chloride, dmso, and dna size standard to perform 50 reactions small or 200 reactions large. Phusion highfidelity dna polymerase phusion hot start ii highfidelity dna polymerase phusion flash highfidelity dna polymerase phusion u hot start dna polymerase phusion u multiplex pcr master mix characteristics blunt or 3a end blunt blunt blunt blunt blunt target length, genomicphage dna. Thermo scientific phusion hot start ii dna polymerase 2 u. Manufactured and qualitycontrolled at new england biolabs, thermo scientific phusion highfidelity dna. As such, conditions recommended below should be used for optimal performance.
Dna used in thermo fisher scientific quality control assays. Incorporating an exciting new technology, phusion dna polymerase brings together a novel pyrococcuslike enzyme with a processivityenhancing domain. Phusion dna polymerase is supplied with standard 5x phusion hf buffer, as well. Due to the high fidelity, even large plasmids can be reliably amplified. Manufactured and qualitycontrolled at new england biolabs, thermo scientific phusion highfidelity dna polymerase offers both high fidelity and robust performance 50x higher fidelity than taq. Robust reactions maximal success with minimal optimization. Article fret image correlation spectroscopy reveals rnapiiindependent ptefb recruitment on chromatin gabriel bidaux,1, corentin le nezet,1 mariano gonzalez pis. The extreme fidelity makes phusion dna polymerase a superior choice for cloning. Phusion dna polymerases thermo fisher scientific us.
The optimal amount of enzyme depends on the amount of template and the length of the pcr product. This is the difference between phusion and taq polymerase. Phusion highfidelity dna polymerase offers extreme performance for all major pcr applications. L reaction depending on the amplicon length and difficulty. Usually 1 unit of phusion hot start ii dna polymerase per 50. With an error rate 50fold lower than that of taq dna polymerase and 6fold lower than that of pyrococcus furiosus dna polymerase 1, phusion is one of the most accurate thermostable polymerases available. It combines the phusion dna polymerase and a reversibly bound, specific affibody ligand. Two master mix formulations are available with hf buffer f531s and f531l and with gc buffer f532s and f532l. Thermo scientificusa 18500 high fidelity dna polymerase packing.
Phusion highfidelity dna polymerases are composed of a dna binding domain fused to a pyrococcuslike proofreading polymerase. Phusion dna polymerase is purified free of contaminating endo and. Thermo scientific phusion hot start ii highfidelity dna. Its unique structure, a novel pyrococcuslike enzyme fused with a processivityenhancing domain, increases fidelity and speed. Thermostable phusion dna polymerase is isolated and purified from an li strain carrying a plasmid with the cloned phusion dna polymerase gene. Datasheet for phusion highfidelity dna polymerase neb. Due to the nature of the phusion highfidelity dna polymerase, the optimal reaction conditions may differ from pcr protocols for standard dna polymerases.
Difference between phusion and taq polymerase compare. Pcr protocol for phusion highfidelity dna polymerase m0530. Due to a proprietary mutation in the so called dutp binding pocket of phusion, phusion u overcomes an important limitation of proofreading enzymes it is able to incorporate dutp and read through uracil present in dna templates. Phusion hot start dna polymerase tends to work better at elevated denaturation and. This kit uses the highly processive thermo scientific phusion hot start ii highfidelity dna polymerase for exponential pcr amplification of dsdna plasmid to. Phusionhigh fidelity dna polymerase possesses the following activities. All that is required is the addition of template, primers and water. Critical optimization parameters for pfu dna polymerase based pcr. Thermo scientific phusion green highfidelity dna polymerase. Phusion highfidelity dna polymerase thermo fisher scientific. The phusion dna polymerase generates long templates with an accuracy and speed previously. As a result of this unique functional pairing, phusion dna polymerases are capable of generating pcr products with very high accuracy and speed.
Phusion highfidelity dna polymerase is supplied with 5x phusion hf buffer, 5x phusion gc buffer, dmso and 50 mm mgcl2 solution. A separate tube of dmso is provided with the product. High fidelity dna polymerase 100 u 20 no phusion high fidelity dna polymerase packing. Solar bio 1950 15 ipgt 5 g 2 no ipgt dioxane free packing. Thermo scientific phusion sitedirected mutagenesis kit, point mutations, insertions and deletions can be introduced in any type of plasmid dna. Thermo scientific phusion highfidelity dna polymerase 2. The phusion dna polymerase generates long templates with an accuracy. Phusion highfidelity 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 mgcl 2. Incorporating an exciting technology, phusion dna polymerase brings together a novel pyrococcuslike enzyme with a processivityenhancing domain.
Both phusion hf buffer and phusion gc buffer provide 1. Finnzymes phusion highfidelity dna polymerase offers extreme performance for all pcr applications. Product information thermo scientific phusion site. High fidelity dna polymerases are important for applications in which the dna sequence needs to be correct after amplification. However, taq polymerase is still used as the standard polymerase enzyme in pcr.
Thermo scientific phusion highfidelity pcr kit wgc. Pcr protocol for pfuultra highfidelity dna polymerase. Phusion sitedirected mutagenesis kit user guide pub. Using a lacibased method modified from previous studies1. Hot start modification of the polymerase prevents amplification of nonspecific products and unwanted degradation of primers prior to first cycle of pcr. Pfuultra highfidelity dna polymerase, part number 600384 conforms to us osha hazard communication 29cfr1910. Since its discovery, several other high fidelity polymerases have been discovered and engineered all with differing processivity, speed and levels of accuracy. Pmc images search for phusion highfidelity dna polymerase figure 1 engineered proofreading dna polymerases have good performance. In the quality control tests of phusion dna polymerase, 7. With the highest fidelity amplification available 280 times higher than taq, q5 dna polymerase results in ultralow error rates, even lower than phusion.
Pfu dna polymerase 10x reaction buffer with mgsom776a. Affibody inhibits the dna polymerase activity at room temperature and thus prevents the amplification of nonspecific products. We enable science by offering product choice, services, process excellence and our people make it. L reaction volume gives good results, but the optimal amount can range from 0. The unique structure and characteristics of phusion dna polymerase make it a superior choice for cloning. Phusion dna polymerases produce blunt end dna products. The phusion highfidelity dna polymerase should be pipetted carefully and gently as the high glycerol content 50% in the storage buffer may otherwise lead to pipetting errors. Phusion highfidelity dna polymerase high performance for.
Thermo scientific phusion highfidelity dna polymerase. Current approaches to identify cell populations that have been modified with deaminase base editing technologies are inefficient and rely on downstre. Phusion hot start ii dna polymerase combines the dna polymerase and a reversibly bound, specific affibody protein. This powerful polymerase mixture is designed to generate pcr products of high yield, high fidelity and high specificity from all types of dna 1. S7 fusion polymerase is a highfidelity dna polymerase that offers extraordinary performance in pcr applications. Please note that protocols with phusion dna polymerase may differ from protocols with other standard polymerases. Phusion hot start ii high fidelity dna polymerase minimizes unwanted secondary mutations. In addition, phusion dna polymerases are tolerant of various inhibitors.
Protocols optimized for phusion dna polymerase can directly be applied to phusion hot start dna polymerase reactions. Phusion hot start ii dna polymerase ensures high fidelity for the exponential amplification, thus minimizing unwanted secondary mutations. Phusion highfidelity dna polymerase is comprised of a dna binding domain fused to a pyrococcuslike proofreading polymerase. L of the 2x master mix depending on the final reaction volume. Phusion highfidelity pcr master mix is convenient 2x mix containing phusion dna polymerase, nucleotides and optimized reaction buffer including mgcl 2. Taq dna polymerase discovery leads to the pcr invention. The concentration of phusion dna polymerase in the phusion pcr master mix has been optimized for best results under a wide range of conditions. Thermo scientific highfidelity dna polymerase selection table. However, due to the novel nature of phusion hot start dna polymerase, optimal reaction conditions may differ from standard enzyme protocols. Phusion hot start ii high fidelity dna polymerase is the most accurate hot start dna polymerase on the market. Q5 highfidelity dna polymerase new england biolabs. The following guidelines are provided to ensure successful pcr using phusion dna polymerase.
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