2 replies to this topic

[Brianlara]

Hi,

    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.

[Trof]

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.

[Brianlara]

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