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

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).