protein

Recombinant Pfu DNA Polymerase

MBS142406

100 Units

205 USD

Recombinant Protein

Recombinant Pfu DNA Polymerase

Pfu DNA Polymerase

Pfu DNA Polymerase; Pfu-DNA Polymerase Recombinant; DNA polymerase; EC 2.7.7.7; Pfu polymerase; Pfu-DNA Polymerase; Pfu DNA Polymerase

DNA-directed DNA polymerase; DNA polymerase; DNA-directed DNA polymerase; DNA polymerase B; Pfu polymerase; Pol I

PF0212; pol

DPOL_PYRFU

E Coli

775

50mM Tris-HCl, pH 8.2, 1mM DTT, 0.1mM EDTA, 0.05% CHAPS and 50% glycerol. Sterile liquid formulation.

Pfu DNA Polymerase although stable at 10 degree C for 5 days, should be stored below -18 degree C.Please prevent freeze-thaw cycles.

1. Ideal for high-fidelity amplification. 2. 3'-5' exonuclease activity provides a low error rate. 3. One of the most thermostable DNA polymerases known. 4. Lack of extendase activity means no unwanted 3' overhangs. 5. Optimal for blunt-end PCR cloning. 6. Optimum temperature near 75 degree C. 7. 95% active after 1-hour incubation at 98 degree C.

10X Buffer with MgSO4: 200mM Tris-HCl (pH 8.8 at 25 degree C), 100mM (NH4)2SO4, 100mM KC1, 1% Triton X-100, 1mg/ml BSA, 20mM MgSO4. PCR Protocol: Add the following components to a amplify 1kb DNA template: 5 ul 10x buffer with MgSO4. 4 ul 2.5mM dNTPs. 1 ul Primers mix (10 uM each). 0.2 ul Pfu-DNA Polymerase. and 38 ul Water. Amplify using the following cycling parameters: Heat Soak: 1 cycle at 94 degree C/4 min. Denaturation: 30 cycles at 94 degree C/30 sec. Annealing: 30 cycles at 58 degree C /30 sec. Extension: 30 cycles at 72 degree C /90 sec. Final: 1 cycle at 72 degree C /5 min. Unit Defenition: 1U of enzyme catalyzes the incorporation of 10nmol of dNTP into acid-insoluble product in 30 minutes at 75 degree C.

ENZYMES; Enzymes; DNA Polymerase

Description: Pfu DNA Polymerase is a thermo-stable enzyme having a Mw of about 90kDa. Pfu DNA Polymerase is derived from E Coli that and cloned from Pyrococcus furiosus strain Vc1 DSM3638. Pfu DNA Polymerase replicates DNA at 75 degree C, catalyzing the polymerization of nucleotides into duplex DNA in the 5 ´ to 3 ´ direction in the existence of magnesium. Pfu DNA Polymerase possesses 3 ´ to 5 ´ exonuclease (proofreading) activity. Base misinsertions that take place during polymerization are swiftly removed by the proofreading activity of the polymerase. Therefore, Pfu DNA Polymerase is suggested for use in PCR and primer extension reactions that require high-fidelity synthesis. Pfu DNA Polymerase-generated PCR fragments are blunt-ended. Introduction: Pfu DNA polymeraseenzyme is found in the hyperthermophilic archaeonPyrococcus furiosus, where it functions in vivoto replicate the organism's DNA. In vitro, Pfu is used to swiftly amplify DNAin the Polymerase Chain Reaction, where the enzyme serves the central function of copying a new strand of DNA during each extension step. Pfu DNA polymerase has superior thermostability and 'proofreading' properties compared to other thermostable polymerases. Unlike Taq DNA polymerase, Pfu DNA polymerase possesses 3' to 5' exonuclease proof reading activity, meaning that it works its way along the DNA from the 5' endto the 3' endand corrects nucleotidemisin corporation errors. Pfu DNA polymerase-generated PCRfragments will have fewer errors than Taq-generated PCR inserts. As a result, Pfu is more commonly used for molecular cloning of PCR fragments than the historically popular Taq. Pfu DNA polymerase is superior for techniques that require high-fidelity DNA synthesis, but can also be used in conjunction with Taq polymerase to obtain the fidelity of Pfu with the speed of Taq polymerase activity.

18976584

NP_577941.1

NC_003413.1

P61875

90,113 Da

Metabolic Pathways (132066); Purine Metabolism Pathway (7520); Purine Metabolism Pathway (307); Pyrimidine Metabolism Pathway (7521); Pyrimidine Metabolism Pathway (309)

100 Units

205 USD

500 Units

290

2500 Units

965