I have two cDNAs; one cDNA is in a pCMV-sport6 and the other in a pCMV5 vector. I wish to begin a stable cell line with either HEK293 cells or HeLa cells. However, both vectors do not have a Neo gene for G418 resistance, a PAC gene for puromycin resistance and/ or a Tet-on for tetracycline resistance. I have two questions:
1. Does this mean I can't produce stable cell lines using these vectors?
2. I only have a pcmv6 vector and a pcdna3.1+zeo vector. Which of these vectors would be better to clone my cDNA into for maintaining a stable cell line?
In our lab we have pcDNA3.1+ vector for stable expression studies. still we are maintaining with our gene of interest. In this vector we inserted GFP downstream of CMV promoter.
I thing you will use pcdna3.1+zeo vector... for your stable expression studies.
Zeocin Selection Antibiotic to develope your stable cells.
http://tools.lifetechnologies.com/content/sfs/manuals/pcdna3_1zeo_man.pdf
Exactly, you can't produce a stable cell line by using these vectors.
Try with pcdna3.1+zeo, it's a very efficient vector. And combined with zeocin in two o three weeks you must have stable colonies.
Will the PCMV6 vector work too? It has a Neo gene and I already have G418 in our lab.
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