Pfu dna polymerase, a lesser proportion of taq dna polymerase, and thermostable dutpase to minimize du dna formation and enhance pfus contribution to blend performance and. Pfu polymerase comparison data we repeatedly compare our pfu polymerase sideby. Pfu dna polymerase exhibits approximately 10 times higher accuracy than taq dna polymerase and amplifies targets up to 34 kb in size. Different to taqdnapolymerase, the pfudna polymerase additionally has a 3 5exonukleaseactivity, the so called. Pfu dna polymerase 3 introduction pfu dna polymerase, a proofreading dna polymerase isolated from pyrococcus furiosus, is an ideal choice for a variety of techniques requiring highfidelity dna synthesis by the polymerase chain reaction pcr. Pfu dna polymerase also exhibits 35 exonuclease proofreading activity, that enables the polymerase to correct nucleotide incorporation errors. The enzyme catalyzes the templatedependent polymerization of nucleotides into duplex dna in the 53 direction. During this process, dna polymerase reads the existing dna strands to create two new strands that match the existing ones. Length of amplification products primestar gxl dna polymerase enables amplification of long dna fragments that cannot be obtained using other commercially available high fidelity pcr enzymes. Base misincorporations that may occur during polymerization are. Products e2016 pfu dna polymerase 1,000 units, 10x reaction buffer with mgso 4 d3001 10 mm dntp mixture 1. One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nanomoles.
To estimate the activity, create a dilution series of purified polymerase in water ranging from 1. It also destroyed the polymerase each time so that fresh enzyme had to be added just after each denaturation step. Pfu dna polymerase is an enzyme found in the hyperthermophilic archaeon pyrococcus furiosus, where it functions to copy the organisms dna during cell division. Pfu dna polymerase is a highly thermostable dna polymerase from hyperthermophilic archaeum pyrococcus furiosus. Pdf a mutant pfu dna polymerase designed for advanced. Dna sequencing with thermus aquaticus dna polymerase and direct sequencing of polymerase chain reactionamplified dna. Pcr fidelity of pfu dna polymerase and other thermostable dna polymerases. A mutant pfu dna polymerase designed for advanced uracil. Phusion highfidelity dna polymerase high performance for. Dna polymerase pfu dna polymerase pyrococcus furiosus modified extremely thermostable proofreading dna poly merase blend, formulated for efficient sitedirected mutagenesis and synthesis of ultra wide range of dna products up to 20 kb in length. It catalyzes the polymerization of nucleotides into duplex dna in the 5. Pfuturbo dna polymerase h320 causes eye irritation. This polymerase derives from the hyperthermophilic.
During the essential dna denaturation step, 94 o c or 95 o c for up to a minute, the dna target was rendered single stranded. Phusion dna polymerase is supplied with standard 5x phusion hf buffer, as well as 5x phusion gc buffer, which can be used for complex or gcrich templates. Pfu dna polymerase stratagene also amplifies dimer templates efficiently, but vent dna polymerase cannot be employed. Description thermo scientific pfu dna polymerase is a highly thermostable dna polymerase from the hyperthermophilic archaeum pyrococcus furiosus. A rapid and efficient onetube pcrbased mutagenesis. Dna polymerases of the time, klenow or t4 dna polymerase. Due to the nature of the phusion highfidelity dna polymerase, the optimal reaction conditions may differ from pcr protocols for standard dna polymerases. Phusion hot start ii green master mix a and proofreading hotstart dna polymerases from other suppliers b merck kod hot start, c agilent pfu ultra ii hs, d kapa biosystems kapa hifi hs, and e takara primestar max were used to amplify a 2 kb fragment from human genomic dna. The enzyme catalyzes the templatedependent polymerization of nucleotides into duplex dna in added. The extension rate of pfu dna polymerase is 1 kb2 minutes. Product information pfu dna polymerase, recombinant pfu. Sehr thermostabile dna polymerase mit korrekturleseaktivitat. Pfu dna polymerase is a highly thermostable dna polymerase from the hyperthermophilic archaeum pyrococcus furiosus. Pfu dna polymerase, native recommendations to lower the.
Size e111801 100 units e111802 500 units e111803 2 500 units. The enzyme catalyzes the templatedependent polymerization of nucleotides into duplex dna in the 5 3 direction. Phusion dna polymerases thermo fisher scientific us. These enzymes are essential for dna replication and usually work in pairs to create two identical dna strands from a single original dna molecule. Pcr fidelity of pfu dna polymerase and other thermostable. Pfu dna polymerase is a highfidelity thermostable dna polymerase with 35 exonuclease activity. Pfu dna polymerase product information 9pim774 promega. The enzyme catalyzes the templatedependent polymerization of nucleotides into duplex dna in 53 direction. A mutant pfu dna polymerase designed for advanced uracilexcision dna engineering article pdf available in bmc biotechnology 101. Pfuturbo dna polymerase warning 10x cloned pfu reaction bufferwarning hazard statements. Pcr amplification reactions, using 10 x pfu buffer with 0.
Thermo scientific pfu dna polymerase is a highly thermostable dna polymerase from the hyperthermophilic archaeum pyrococcus furiosus. Pfu dna polymerase replicates dna at 75c, catalyzing the polymerization of nucleotides in the 5. The fidelity is more than tenfold higher than that of taq. Pfu dna polymerase remains 95% active after 2 hours incubation at 95c.
Pfu dna polymerase was selected for this method because it is reported to have 12fold increased fidelity of dna synthesis compared to taq polymerase 1. To date, the relative performance of dna polymerase blends, proofreading pcr enzymes, and hot start. The pcr amplicon was subsequently sanger sequenced. Pfu dna polymerase, derived from the hyperthermophilic archae pyrococcus furiosus, has superior thermostability and proofreading properties compared to the other thermostable polymerase. Pfu dna polymerase is a thermostable enzyme of approximately 92kda isolated from pyrococcus furiosusdsm3638 5. In 1999, the crystal structure of the dna polymerase tgo from the archea thermococcus gorgonarius was solved, revealing the nature of the uracilbinding pocket, and allowing the design of mutant tgo and pfu polymerases with reduced stalling at uracilcontaining dna. Pfu dna polymerase 500 units 90035001 2500 units 900325001 10x reaction buffer 2 ml 90035002 2 ml x 5 900325002 concentration. Pfu dna polymerase is a highfidelity, thermostable enzyme of approximately 90kda isolated from pyrococcus furiosus. Thermo scientific phusion highfidelity dna polymerase was created by fusing a dsdnabinding domain to a pyrococcuslike proofreading polymerase. The template for this assay is the 6hispfusso7dpet28 plasmid encoding the polymerase. 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. Highfidelity pcr enzymes are used for applications requiring high accuracy during dna amplification such as cloning, sequencing or mutagenesis.
Pfu dna polymerase is recommended for use in pcr and primer extension reactions that require high fidelity 14. Pfudna polymerase is a thermostable enzyme that replicates dna at 75c. Pfu dna polymerase is a thermostable enzyme that replicates dna at 75c. Pfu dna polymerase an overview sciencedirect topics. The processivity of phusion dna polymerases is 10fold greater than that of pfu dna polymerase and twice favored over most other polymerases for accuracy, speed, and value that of taq dna polymerase. Dna polymerase is an enzyme that synthesizes dna molecules from deoxyribonucleotides, the building blocks of dna. Product information pfu dna polymerase, recombinant.