Thermo Scientific™ Phusion High-Fidelity PCR Master Mix with HF or GC Buffer

Description

Thermo Scientific Phusion High-Fidelity DNA polymerases set a gold standard for high performance PCR. Featuring an error rate 50-fold lower than that of Taq and 6-fold lower than that of Pfu, Phusion High-Fidelity DNA Polymerase is excellent choice for cloning and other applications requiring high fidelity. Phusion DNA polymerases offer robust performance with short protocol times, even in the presence of PCR inhibitors, and generate higher yields with lower enzyme amounts than other DNA polymerase.

Phusion High-Fidelity PCR Master Mix is convenient 2X mix containing Phusion DNA Polymerase, nucleotides, and optimized reaction buffer including MgCl₂. Two master mix formulations are available: with HF Buffer (F-531S and F-531L) and with GC Buffer (F-532S and F-532L). The error rate of Phusion DNA Polymerase in HF Buffer (4.4 x 10^-7) is lower than that in GC Buffer (9.5 x 10^-7). Therefore, the master mix with HF Buffer should be used as the default master mix for high-fidelity amplification. However, GC Buffer can improve the performance of Phusion DNA Polymerase on some difficult or long templates, such as GC-rich templates or those with complex secondary structures.

Highlights

• High fidelity (25X Taq)
• Fast PCR due to short extension times (15–30 s/kb)
• Robust performance, minimal optimization needed

Applications

• Amplification of difficult (GC-rich) templates
• High-fidelity PCR
• Cloning
• Template generation for sequencing
• Long-range PCR (up to 20 kb)
• Mutagenesis
• High throughput PCR

Using Phusion DNA polymerases

Annealing rules for Phusion DNA polymerases are different from many common DNA polymerases (such as Taq DNA polymerases).