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Can the pcDNA5/FRT/TO vector be used for transient overexpression?

transfection

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#1 Norbert

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Posted 07 July 2014 - 12:15 PM

Hi,

 

I have my gene of interest in pcDNA5FRT/TO and would like to generate a stable cell line. Before generating the stable line, however, I would like to test expression by transient transfection.

 

Can the pcDNA5FRT/TO vector be used for transient transfection? If so, do I need to add doxycycline to induce expression (as would be done with a stable line ...)? In a couple of papers that I found, addition of doxycyclin during transient overexpression of a construct in pcDNA5/FRT/TO was not specifically mentioned.

 

Thank you,

 

Norbert



#2 bob1

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Posted 07 July 2014 - 12:57 PM

Yes you do need to add dox.  The FRT/TO plasmid has two copies of the tet repressor inserted inside the promoter region, which repress transcription unless Dox is present.







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