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  • FAQ: What if my primer extension reaction yields no product or a smear?

    * Follow recommended conditions, included in product literature
    * Optimize annealing and extension temperatures
    * Optimize Mg2+ level
    * Optimize amount of polymerase
    * Purify DNA template by phenol/chloroform extraction and alcohol precipitation
    * Avoid high salt carry-over into primer extension reaction
    * Make sure dNTP solution has not undergone hydrolysis - try fresh dNTPs
    * Cosolvents such as 100 mg/ml BSA, 1-10% formamide or 1-10% DMSO may help with some primer: templates