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