Pfu DNA Polymerase: A High-Fidelity Thermostable Enzyme for Accurate DNA Amplification  

Pfu DNA Polymerase: A High-Fidelity Enzyme for PCR

Pfu DNA Polymerase is a high-fidelity, thermostable enzyme isolated from the hyperthermophilic archaeon Pyrococcus furiosus. It is commonly used in polymerase chain reaction (PCR) applications due to its ability to synthesize DNA with high accuracy and its stability at high temperatures. Pfu DNA Polymerase has a 3'→5' exonuclease activity, also known as proofreading activity, which allows it to remove incorrectly incorporated nucleotides and thus reduce the error rate of DNA synthesis.

Key Features of Pfu DNA Polymerase

• High fidelity: Pfu DNA Polymerase has an error rate of approximately 2.5 x 10^-5 errors per base pair, making it one of the most accurate DNA polymerases available.
• Thermostability: Pfu DNA Polymerase is stable at high temperatures, with an optimal reaction temperature of 75°C. This makes it ideal for PCR applications, as it can withstand the repeated heating and cooling cycles required for DNA amplification.
• 3'→5' exonuclease activity: Pfu DNA Polymerase has proofreading activity, which allows it to remove incorrectly incorporated nucleotides and thus reduce the error rate of DNA synthesis.

Applications of Pfu DNA Polymerase

Pfu DNA Polymerase is widely used in a variety of molecular biology applications, including:

• PCR: Pfu DNA Polymerase is commonly used for high-fidelity PCR applications, where accurate DNA synthesis is critical.
• Site-directed mutagenesis: Pfu DNA Polymerase is used in site-directed mutagenesis, a technique used to introduce specific mutations into DNA sequences.
• DNA sequencing: Pfu DNA Polymerase is used in some DNA sequencing methods, such as Sanger sequencing.

Advantages of Pfu DNA Polymerase

Pfu DNA Polymerase offers several advantages over other DNA polymerases, including:

• High fidelity: Pfu DNA Polymerase has one of the lowest error rates of any DNA polymerase, making it ideal for applications where accurate DNA synthesis is critical.
• Thermostability: Pfu DNA Polymerase is stable at high temperatures, making it suitable for use in PCR applications.
• Proofreading activity: Pfu DNA Polymerase has proofreading activity, which further reduces the error rate of DNA synthesis.

Disadvantages of Pfu DNA Polymerase

Pfu DNA Polymerase also has some disadvantages, including:

• High cost: Pfu DNA Polymerase is more expensive than some other DNA polymerases, such as Taq DNA Polymerase.
• Slow processivity: Pfu DNA Polymerase is relatively slow compared to some other DNA polymerases, such as Taq DNA Polymerase.

Overall, Pfu DNA Polymerase is a powerful tool for molecular biology research. Its high fidelity, thermostability, and proofreading activity make it ideal for a variety of applications, including PCR, site-directed mutagenesis, and DNA sequencing.

Product Introduction

Pfu DNA Polymerase is a high-fidelity, thermostable DNA polymerase isolated from the hyperthermophilic archaeon Pyrococcus furiosus and recombinantly expressed in Escherichia coli. It exhibits exceptional amplification efficiency and fidelity, making it an invaluable tool for a wide range of molecular biology applications.

Key Features

• Superior Amplification Efficiency: Pfu DNA Polymerase boasts an impressive amplification rate of 1 kb/10 seconds, significantly outperforming conventional DNA polymerases like Taq DNA Polymerase.
• Exceptional Fidelity: Pfu DNA Polymerase excels in maintaining the integrity of DNA sequences, boasting an error rate of approximately 2.5 x 10^-5 errors per base pair. This remarkable fidelity ensures the accuracy of PCR amplification, making it ideal for sensitive applications.
• Robust Long Fragment Amplification: Pfu DNA Polymerase demonstrates exceptional ability to amplify long DNA fragments, effectively amplifying sequences up to 10 kb in length. This capability makes it well-suited for applications such as long-PCR and cDNA synthesis.

Applications

Pfu DNA Polymerase finds widespread application in various molecular biology techniques, including:

• High-Fidelity PCR: Pfu DNA Polymerase is the preferred enzyme for high-fidelity PCR applications where accurate DNA amplification is crucial. Its superior fidelity ensures the integrity of amplified sequences, making it ideal for cloning, mutagenesis, and gene expression studies.
• Long-PCR: Pfu DNA Polymerase's ability to amplify long DNA fragments makes it an indispensable tool for long-PCR applications. It effectively amplifies sequences up to 10 kb in length, enabling researchers to study large genes and genomic regions.
• cDNA Synthesis: Pfu DNA Polymerase is widely used for cDNA synthesis, the process of converting RNA into complementary DNA (cDNA). Its high fidelity ensures the accuracy of cDNA synthesis, allowing for reliable downstream applications such as gene expression analysis.

In summary, Pfu DNA Polymerase stands as a powerful tool for molecular biology research, offering exceptional amplification efficiency, fidelity, and long fragment amplification capabilities. Its versatility and accuracy make it a valuable asset for a wide range of applications, including high-fidelity PCR, long-PCR, and cDNA synthesis.

Product Description

Pfu DNA Polymerase is a high-fidelity, thermostable DNA polymerase isolated from the hyperthermophilic archaeon Pyrococcus furiosus and recombinantly expressed in Escherichia coli. It demonstrates exceptional amplification efficiency and fidelity, making it an invaluable tool for a wide range of molecular biology applications.

Key Features

• Superior Amplification Efficiency: Pfu DNA Polymerase boasts an impressive amplification rate of 1 kb/10 seconds, significantly outperforming conventional DNA polymerases like Taq DNA Polymerase.
• Exceptional Fidelity: Pfu DNA Polymerase excels in maintaining the integrity of DNA sequences, boasting an error rate of approximately 2.5 x 10^-5 errors per base pair. This remarkable fidelity ensures the accuracy of PCR amplification, making it ideal for sensitive applications.
• Blunt-Ended Amplification Products: Pfu DNA Polymerase generates blunt-ended products in the amplification reaction, simplifying downstream applications such as cloning and ligation.

Applications

Pfu DNA Polymerase finds widespread application in various molecular biology techniques, including:

• High-Fidelity PCR: Pfu DNA Polymerase is the preferred enzyme for high-fidelity PCR applications where accurate DNA amplification is crucial. Its superior fidelity ensures the integrity of amplified sequences, making it ideal for cloning, mutagenesis, and gene expression studies.
• Blunt-End Ligation: The blunt-ended products generated by Pfu DNA Polymerase facilitate seamless ligation into blunt-end cloning vectors, streamlining the cloning process.
• Site-Directed Mutagenesis: Pfu DNA Polymerase's high fidelity makes it an ideal choice for site-directed mutagenesis, a technique used to introduce specific mutations into DNA sequences.

In summary, Pfu DNA Polymerase stands as a powerful tool for molecular biology research, offering exceptional amplification efficiency, fidelity, and blunt-end amplification capabilities. Its versatility and accuracy make it a valuable asset for a wide range of applications, including high-fidelity PCR, blunt-end ligation, and site-directed mutagenesis.