Overview:

• High-fidelity thermostable DNA polymerase for error-free polymerase chain reaction.
• Possess 3′ to 5′ exonuclease activity for proofreading activity
• Produces blunt-end DNA fragments

Description

• Phusion DNA polymerase is a high-fidelity, thermostable DNA-dependent DNA polymerase engineered for accurate and efficient DNA amplification, especially in PCR applications requiring precision and speed. It is a fusion enzyme, combining a thermophilic DNA polymerase with a processivity-enhancing DNA-binding domain derived from a Pyrococcus-like archaeal polymerase and an accessory protein. This fusion significantly increases the enzyme’s processivity, stability, and fidelity compared to standard polymerases like Taq.
• Phusion exhibits robust 5′→3′ polymerase activity and possesses an intrinsic 3′→5′ exonuclease (proofreading) activity, which allows it to correct misincorporated nucleotides during DNA synthesis. As a result, it has an exceptionally low error rate—approximately 50 times lower than Taq polymerase—making it ideal for applications requiring high sequence accuracy, such as cloning, mutagenesis, next-generation sequencing library preparation, and functional genomics.
• Optimally active at 68–72°C, Phusion polymerase is capable of amplifying long DNA targets (up to ~40 kb for genomic DNA and ~20 kb for cDNA), and its high processivity results in shorter extension times—often 15–30 seconds per kb. It also performs well under fast PCR cycling conditions and in the presence of challenging templates with high GC content or secondary structures.
• Phusion polymerase is supplied with optimized buffers that support hot-start capability and minimize nonspecific amplification. The enzyme’s fidelity and performance make it a preferred choice in both academic and industrial settings where sequence accuracy and efficiency are critical.
• In summary, Phusion DNA polymerase is a high-performance, proofreading enzyme designed for accurate, high-yield DNA amplification across a range of molecular biology applications. Its superior fidelity and robustness make it an excellent tool for cloning, diagnostics, and high-throughput DNA synthesis.

REFERENCES

• Wang et al., 2004. A novel strategy to engineer DNA polymerases for enhanced processivity and improved performance in vitro. Nucleic Acids Res. 32(3), 1197-1207. PMID-14973201; Full-Text Links: academic.oup (Download PDF), PMC373405 (Download PDF)