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PCR products form with DNA, but not with c-DNA

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#1 Brianlara



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Posted 12 August 2011 - 02:15 PM


I am currently trying to test gene expression of a transgene (inserted in poplar) using q-PCR. I designed primers and tested them first on DNA and obtained bands on an agarose gel (got bands for the positive control as well). Then, I tried testing the primers on c-DNA. While I got bands for ubiquitin, I did not get any bands for my primers. What could be the possible reasons? Any advice on how to proceed would be appreciated. I have the gel pictures and can upload them if required.

Thanks in advance.

#2 Trof


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Posted 14 August 2011 - 04:27 AM

How do you designed your primers? They're obviously not intron spanning, but are you sure they're not located in the intron part of a gene? Or your transgene doesn't contain introns?
Other reason may be, that your transgene is simply not expressed.

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#3 Brianlara



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Posted 16 August 2011 - 02:59 PM

Thanks for your reply. I checked; the primers don't seem to be located in the intron part.

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