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  • FAQ: I am having trouble amplifying a template that is longer than 5kb. How can I optimize my product yield using Phusion® High-Fidelity DNA Polymerase?

    * Use more template. Sample concentration may be too low.
    * Template DNA may be damaged. Use carefully purified template.
    * Optimize enzyme concentration by testing a titration of enzyme in the reaction (0.25-2 units/50μl reactions)
    * Increase number of cycles.
    * If HF buffer has failed, try using GC buffer. GC buffer allows better amplification of GC rich and longer templates, but slightly lowers fidelity.
    * Lengthen extension time.