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HEK293T and G418


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#1 science noob

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Excellent

Posted 18 September 2012 - 07:08 PM

Been culturing HEK293T with various concentrations of G418 to optimise the right G418 concentration for transfection (my plasmid has a Neomycin resistance insert in it).
It seems like the cells are pretty happy sitting in G418 up to around 4mg/ml. Is that considered as a high concentration? According to data sheets, IC50 of G418 is around 2mg/ml but most papers/labs use 0.5-1mg/ml.

Seen posts about HEK293T being already resistant to G418 as opposed to HEK293. Can someone clarify/confirm this??

I do know that HEK293FT is G418-resistant.

#2 cellthetruth

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Neutral

Posted 18 September 2012 - 11:12 PM

Hi,

yes, HEK293T cells are G418 (Geneticin) resistant. They are stable transfectants with an aph3' resistance cassette. So you can not select for your plasmid. I would suggest that you replace your NeoR with a Hygromycin resistance cassette. Usual concentration we use is 200 ug/ml for selection and 100 ug/ml for maintaining the cells.




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