Our lab is working on developing the stable gene expressing clone in DG44 cells,. After transfection, transient gene expression is working, after growing the cells in the media with out GHT and addition of methotrexate (20nM), we donot see any stable gene expression (SGE) in the transfected cells. How address this problem of SGE in DG44 cells.
Why methotrexate? Which is the antibiotic of selection of your plasmid?
Hello Rafael ,
Thanks for the reply and suggestion, we have puromycin selection marker will try the selection marker.
Hello Alex,
Thanks for the link to the paper, which is a parallel work, will be of good use. Will try to replicate the work.
regards,
Hello,
You can use both MTX and puromycin for selecting the transfected CHO- DG44 cells. If possible use CHO-S cells instead of CHO-DG44 cells for SGE.
Thanks!
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