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I have run a gradient PCR.. and band 6 after the ladder is clearly the brightest. But all bands appear below the ladder. What does that mean? It is extracted mDNA that was loaded.

Thank you in advance!

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Below the ladder means primer dimer. You have no product.

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  • $\begingroup$ Thank you! That explains it. I will try deciding primer amounts and hope for the best! $\endgroup$
    – Brian McGarry
    Commented Nov 10, 2017 at 17:40
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    $\begingroup$ In my experience, altering primer concentration won't do anything. I'd rethink the annealing temp, maybe try a more robust Taq, double-check the quality of your input DNA, and double-check that you have the right primers, and that they actually should make a product. $\endgroup$
    – swbarnes2
    Commented Nov 10, 2017 at 21:03

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