Two optimized buffers are available for Thermo Scientific Phusion High-Fidelity DNA Polymerases: 5X Phusion HF Buffer and 5X Phusion GC Buffer. The error rate of Phusion DNA Polymerase in HF Buffer (4.4 × 10-7) is lower than that in GC Buffer (9.5 × 10-7). Therefore, the HF Buffer should be used as the default buffer 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. For applications such as microarray or DHPLC, where the DNA templates need to be free of detergents, we recommend using detergent-free reaction buffers.
5X Phusion Green Buffers include a density reagent and two tracking dyes for direct loading of PCR products on a gel. The colored buffers do not interfere with the enzyme performance and are compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.