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  • FAQ: Why are there low molecular weight discrete bands on an agarose gel after a PCR using Phusion® Hot Start Flex DNA Polymerase?

    Non-specific products are being amplified. To avoid this we suggest the following:
    • Re-calculate primer Tms using NEB's Tm Calculator .
    • Raise the annealing temperature.
    • Lower polymerase concentration.
    • Shorten extension time.
    • Optimize Mg2+ concentration.
    • Titrate template amount.
    • Lower primer concentration.