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
- - - - -

Question in Generating double stable cell line


  • Please log in to reply
3 replies to this topic

#1 avitas

avitas

    member

  • Active Members
  • Pip
  • 9 posts
0
Neutral

Posted 06 May 2010 - 08:29 PM

I have a question in generating double stable cell line.
I am trying to generate a stable cell line w/ 2 genes which usually form a heterodimer. I understand that doing double stable cell line I need to use 2 different selection markers. Do people usually do cotransfection of both expression vectors and add 2 different antibotics to do selection at the same time? or Do I need to do sequential transfection, as in generate a stable cell line for one of the genes first, then use that stable cell line and transfect the 2nd gene and select for it the 2nd round?

Thanks in advance for any help!!

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,830 posts
414
Excellent

Posted 09 May 2010 - 04:29 PM

It can be done either way, though the sequential option is probably easier, if somewhat longer.

#3 avitas

avitas

    member

  • Active Members
  • Pip
  • 9 posts
0
Neutral

Posted 11 May 2010 - 10:28 AM

It can be done either way, though the sequential option is probably easier, if somewhat longer.


Thanks for the response.

Any suggestion on which 2 selection markers are easier to use?

#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,830 posts
414
Excellent

Posted 11 May 2010 - 04:11 PM

Geneticin and hygromycin are common enough. Make sure you do a kill curve to see the appropriate dose of the selective agent before you do the transfections.




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

©1999-2013 Protocol Online, All rights reserved.