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Black dots contaminant in stable cell lines - antibiotics

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#1 Lfs



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Posted 28 June 2016 - 09:14 AM



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


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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.

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