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Cell culture of B16OVAs


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#1 Ahrenhase

Ahrenhase

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Posted 03 March 2014 - 12:14 PM

So I just found out that the B16OVA cells I'm using carry the plasmid pAc-neo-OVA, and I was not told to use G418 selection when culturing.  To compound my problems, I mistakingly cultured them in RPMI+10% FBS rather than DMEM+10%FBS, and the cells lost melanin expression in 2 passages. 

 

 

What are the chances I lost my OVA expression in these cells.  There doesn't seem to be an origin that replicates in eukaryotes on this plasmid. 






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