Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

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


  • Please log in to reply
2 replies to this topic

#1 Brianlara

Brianlara

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 12 August 2011 - 02:15 PM

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.

#2 Trof

Trof

    Brain on a stick

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,199 posts
109
Excellent

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.

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon


#3 Brianlara

Brianlara

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

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.




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.