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!
- - - - -

Question: Can I cut out rb glob PA Terminator

cloning dna plasmid restriction enzyme

  • Please log in to reply
1 reply to this topic

#1 Fabian Yu

Fabian Yu


  • Members
  • Pip
  • 1 posts

Posted 21 January 2015 - 09:11 AM

Hi There,


I am doing a cloning project with the pCAGGs promoter.  I already ligated a fragment of my gene of interest  in the plasmid, and now I need to add an IRES::Puro sequence.  So far my ligation is not working but I'm not sure whether it is b/c I cut out the rb glob PA Terminator in the plasmid.  I know the poly a site helps with transcription but is it essential for my plasmid? Only reason I did that was due to the cut site  issue 








  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 140 posts

Posted 21 January 2015 - 06:23 PM

Looks like you are making a bicistronic vector: gene of your interest (with stop codon but without polyA signal)-IRES-puro-polyA.


If so, you do need polyA at the end of IRES-puro cassette.

So. Now that you have your first ever question on bioforum answered (or not), mail yourself your username and password so you don't forget them, and then come back soon to update us on how it all worked out. That's how you build Karma in science.

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

©1999-2013 Protocol Online, All rights reserved.