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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 PCR Master Mix?

    * Increase template amount. Sample concentration may be too low.
    * Template DNA may be damaged. Use carefully purified template.
    * Lengthen extension time.
    * Increase the number of cycles.
    * Lower annealing temperature.
    * Titrate DMSO (2-8%) in the reaction.
    * Denaturation temperature may be too low. Optimal denaturation temperature for most templates is 98°C or higher.
    * Denaturation time may be too long or too short. Optimize the denaturation time.