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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® Hot Start Flex 2X Master Mix?

    • Use more template. Sample concentration may be too low.
    • Template DNA may be damaged. Use carefully purified template.
    • Increase number of cycles.
    • Lengthen extension time.
    • Re-calculate primer Tms using NEB's Tm Calculator .