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
- - - - -

Black dots contaminant in stable cell lines - antibiotics


  • Please log in to reply
1 reply to this topic

#1 Lfs

Lfs

    member

  • Active Members
  • Pip
  • 28 posts
1
Neutral

Posted 28 June 2016 - 09:14 AM

Hello!

 

I'm facing the "black dot contamination" problem in my stable cell line. The problem is that I cannot throw this culture away so I started to use the antibiotics (Cipro/Piper/Tazo, 10 ug/ml each + Pen/Strep) described in this paper http://www.ncbi.nlm....les/PMC2849847/.

 

(*stable cell lines were made by transfection of a sleeping beauty transposon vector (pSBTet-BP) inducible by doxycycline with a Blue Protein for visualizing Puromycin positive clones).

 

Strangely, after 1-3 days of treatment, most of the blue fluorescence of my positive stable cell line pool started to disappear!

 

Do guys know if Ciprofloxacin and/Piperacillinlin and/or Tazobactam can interfere in gene expression or it just decreasing cell fluorescence for no reason?

 

Thank you in advance!

 



#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,362 posts
509
Excellent

Posted 06 July 2016 - 06:36 AM

Have you been culturing the cells with dox while doing the other treatments? If not, you might be losing the signal by methylation of the inserted gene.






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

©1999-2013 Protocol Online, All rights reserved.