Pfu DNA Polymerase Protein

• Catalog Number: orb90215
• Category: Proteins
• Description: Recombinant Pfu-DNA Polymerase protein
• Form/Appearance: Sterile liquid formulation.
• Source: Escherichia Coli
• Storage: Stability: Pfu DNA Polymerase although stable at 10°C for 5 days, should be stored below -18°C.Please prevent freeze-thaw cycles
• Buffer/Preservatives: 50mM Tris-HCl, pH 8.2, 1mM DTT, 0.1mM EDTA, 0.05% CHAPS and 50% glycerol.
• Alternative names: DNA polymerase, EC 2.7.7.7, Pfu polymerase, Pfu-DN
• Note: For research use only
• Application notes: Applications: 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°C. 7. 95% active after 1-hour incubation at 98°C. Unit Definition: 1U of enzyme catalyzes the incorporation of 10nmol of dNTP into acid-insoluble product in 30 minutes at 75°C. 10X Buffer with MgSO4: 200mM Tris-HCl (pH 8.8 at 25°C), 100mM (NH4)2SO4, 100mM KCl, 1% Triton X-100, 1mg/ml BSA, 20mM MgSO4. PCR Protocol: Add the following components to amplify 1kb DNA template: 0.2µl Pfu-DNA Polymerase.4µl 2.5mM dNTPs.5µl 10x buffer with MgSO4. 1µl Primers mix (10µM each).1µl Template.38µl ddH2O. Amplify using the following cycling parameters: Heat Soak: 1 cycle at 94°C/4 min.Denaturation: 30 cycles at 94°C/30 sec.Annealing: 30 cycles at 55°C /30 sec.Extension: 30 cycles at 72°C /90 sec. Final: 1 cycle at 72°C /5 min
• Expiration Date: 6 months from date of receipt.