Generating double stable cell line
Posted 23 April 2009 - 07:37 AM
I have a problem concerning my cell line. All DNA constructs, prior used with another cell line, use the neomycine resistance gene for generation of stable cell lines. My cell line was immortalized with pE1A-neo plasmid which means that this cell line is already neomycin resistant. Alternatives are e.g. puromycin or hygromycin etc. Who has experiences with selection alternatives to neomycine? Which plasmids are recommended for this attempt?
Posted 23 April 2009 - 08:35 AM
It contains CMV promoter and allows the expression of your gene of interest in fusion with the selection marker (it is a mRNA fusion not a protein fusion).
Thereby you can reduce the screening effort a bit since resistent clones are likely to express your gene of interest as well.
I find puromycin selection also faster and more reliable.
Posted 26 April 2009 - 05:57 PM