I have tranceduced the NK92MI cell line with a puromycin resistance plasmid, and I wanna to select the positive clone, there is anyone who knows the optical puromycin concentration?
Dear Junwe
We havent tried the NK92MI cell line, but when we use selection we normally establish a kill curve of the parent cell line, i.e. determine which concentrations of puromycin will just be efficient enough for killing the parent cells.
BR Birger
1.5-2ug/mL final conc is usually effective on most cell lines (if you don't optimise it as suggested above) - as long as you have a control plate of cells alongside. You should see a significant effect after 3 days, keep changing media and maintain the puro until all the non-transfected cells are dead. Maintain clones in 1 ug/mL.
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