3 replies to this topic

[avitas]

I have a question in generating double stable cell line.
I am trying to generate a stable cell line w/ 2 genes which usually form a heterodimer. I understand that doing double stable cell line I need to use 2 different selection markers. Do people usually do cotransfection of both expression vectors and add 2 different antibotics to do selection at the same time? or Do I need to do sequential transfection, as in generate a stable cell line for one of the genes first, then use that stable cell line and transfect the 2nd gene and select for it the 2nd round?

Thanks in advance for any help!!

[bob1]

It can be done either way, though the sequential option is probably easier, if somewhat longer.

[avitas]

bob1, on May 9 2010, 05:29 PM, said:

It can be done either way, though the sequential option is probably easier, if somewhat longer.

Thanks for the response.

Any suggestion on which 2 selection markers are easier to use?

[bob1]

Geneticin and hygromycin are common enough.  Make sure you do a kill curve to see the appropriate dose of the selective agent before you do the transfections.