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  • Help with PCR

    Good morning,

    I'm student of master and my thesis is with animals virus. I have problems with the PCR: I ruled out contamination of the reactives, but in the positive samples appear a unknown band, the negative control is negative.

    Can somebody help me with papers or handbooks that talk about mistakes in PCR?

    Thanks for all.

  • #2
    Can you give us a little more information, it may be that the unknown band is due to non-specific binding of your primers? What size is it roughly? What cycling conditions did you use, composition of your master mix etc? Bio-rad PCR troubleshooting is always a good place to start: http://www.bio-rad.com/en-uk/applica...roubleshooting

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    • #3
      In your positive samples, do you see your expected band AND an unknown band or you just see unknown/unexpected band? Is it PCR that is known to produce good results or you are trying to develop a PCR ?

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