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
- - - - -

Maxi Prep cDNA contamination


  • Please log in to reply
2 replies to this topic

#1 PCR Novice

PCR Novice

    member

  • Active Members
  • Pip
  • 16 posts
0
Neutral

Posted 01 April 2009 - 07:44 AM

Hi

In our department here we have been setting up a PCR lab, for over a year now, and have run into a major problem with cDNA contamination-it just seemed to keep spreading and spreading no matter what we did, I have been reading numerous blogs here and getting tips and I think we finally have it under control.

The problem I have is that we prepared a Maxi-prep and I seem to have contaminated it with cDNA-we do what we call an 'elegant negative' where we do a PCR and use primers that shoudnt work on the plasmid, it worked a month ago but with the amount of contamination we had we wanted to repeat it before we took the next step. Is there anyway I can clean-up the plasmid without having to redo the Maxi-prep???

Thanks :)

#2 molgen

molgen

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 167 posts
0
Neutral

Posted 02 April 2009 - 03:35 AM

Try running your plasmid on a gel.
Extract it from the gel and then redo the PCR.

#3 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,467 posts
247
Excellent

Posted 02 April 2009 - 06:28 AM

Given the sensitivity of PCR, I think it is unlikely that a gel purificaton will remove template contamination. Perhaps it is worth a try.




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

©1999-2013 Protocol Online, All rights reserved.