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
- - - - -

primer dimer or what?


  • Please log in to reply
2 replies to this topic

#1 Tintin

Tintin

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 31 July 2009 - 05:56 AM

Recently I used a few pair of primers to test genomic DNA contamination in my cDNA (RT from total RNA). The primers are one in nontranscript region, the other in exon. If no genomic DNA contamination, it should has no band, right? But I always have a <75bp bands, even I tried several different pair. And when I do other traditional PCR, if my product has low yield, I have this suspicious band as well.
Is it a primer dimer? How can I avoid it?

#2 pcrman

pcrman

    Epigenetist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,165 posts
68
Excellent

Posted 31 July 2009 - 10:03 PM

If the expected size of your prodcut is > 75 bp, the the 75 bp one is most likely primer dimer. In this case probably you have to redesign your primers because your current primers form self or inter pairing.

#3 dreww

dreww

    RNA Follower

  • Active Members
  • PipPipPipPipPip
  • 36 posts
0
Neutral

Posted 01 August 2009 - 11:03 PM

Yes, more than likely. You can run a Control with no template and compare the bands. Also, if it's genomic DNA, your bands should sit high. If you also know that your target band is >75bp, that should answer your problem.




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

©1999-2013 Protocol Online, All rights reserved.