What is the difference between Pfu polymerase and Taq polymerase?
The main difference between Pfu and alternative enzymes is that Pfu has superior thermostability and proof-reading properties. Unlike Taq DNA polymerase, Pfu DNA polymerase also possesses 3′->5′ exonuclease proofreading activity, resulting in PCR fragments with fewer errors than Taq-generated PCR inserts.
What is Taq and pfu?
Unlike Taq DNA polymerase, Pfu DNA polymerase possesses 3′ to 5′ exonuclease proofreading activity, meaning that as the DNA is assembled from the 5′ end to 3′ end, the exonuclease activity immediately removes nucleotides misincorporated at the 3′ end of the growing DNA strand.
Is PFU faster than Taq?
Unlike Taq DNA polymerase, Pfu DNA polymerase possesses 3′ to 5′ exonuclease proofreading activity, meaning that it works its way along the DNA from the 5′ end to the 3′ end and corrects nucleotide-misincorporation errors. The downside of Pfu is its speed which is slower that that of Taq.
Is PFU used in PCR?
Base misincorporations that may occur during polymerization are rapidly excised by this proofreading activity. Consequently, Pfu DNA Polymerase is recommended for use in PCR and primer extension reactions that require high-fidelity synthesis. Pfu DNA Polymerase-generated PCR fragments are blunt-ended.
Why is Pfu polymerase better than Taq polymerase?
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.
Is PFU a high fidelity polymerase?
Pfu is the most commonly used high fidelity DNA polymerase for PCR and exhibits an average error rate of 1.3 × 10−6 mutations/bp/duplication [12], which is approximately eight-fold more accurate than Taq DNA polymerase.
Is Taq polymerase High Fidelity?
One unit of Platinum® Taq DNA Polymerase, High Fidelity, is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into DNA in 30 min at 74°C.
Is Pfu a high fidelity polymerase?
Why PFU is more efficient than the Taq polymerase?
Why are Vent polymerase and Pfu more efficient than the Taq polymerase?
7. Why are vent polymerase and Pfu more efficient than the Taq polymerase? Sol:(a) Because of proofreading activity.
What should the temperature be for pfu polymerase?
Extension The optimal extension temperature for Pfu DNA Polymerase is 70-75°C. The recommended extension step is 2 min/kb at 72°C for PCR products up to 2 kb. For larger products, the extension time should be prolonged by 1 min/kb.
How is Taq DNA polymerase used in PCR?
In addition, Taq DNA Polymerase exhibits deoxynucleotidyl transferase activity, which frequently results in the addition of extra adenines at the 3′-end of PCR products. Recombinant Taq DNA Polymerase is the ideal tool for standard PCR of templates 5 kb or shorter.
When to use 10x Taq buffer without detergent?
Accuracy is an inverse of the error rate and shows an average number of correct nucleotides incorporated before an error occurs. • The 10X Taq Buffer without Detergent is recommended for microarray experiments. For Research Use Only. Not for use in diagnostic procedures.