Which antibiotic to choose? - puro vs hygro vs zeo etc (Jan/11/2007 )
Hi,
There are many antibiotic markers available for stable transfection and I was wondering how to chose one over another. I've heard that puro is the best in terms of efficiency and speed (from both my PI and Fred_33 in these forums), but haven't been able to find any literature on this, nor have I been able to find (peer-reviewed) literature on the other available markers. I think that I would like to use puro, except it is difficult to find a vector suited to my needs with a puro R gene. Instead, I'm looking at another vector available with Neo, Zeo or Hygro R genes (Invitrogen pcDNA3.1/).
Any help would be most appreciated!
Dave
Neomycine selection is useful, as the same resistence gene will give bacteria resistence to kanamycine. Of course you will need prokaryote promoters for it to work in bacteria, but there are vectors that have that.
I haven't used zeocine lately, and my books aren't with me. All I remember is that this marker didn't really do for me. But that was due to the plasmid structure I was using.
Hygromycine... is a bit expensive, and require quite high concentration to work. It doesn't give completely clean selection. However don't look totally down on this. If your resistence gene is not working well, a weaker selection agent will allow you to see colonies carrying your plasmid, where a stronger selection agent would wipe out all colonies.
There is blasticidine as well. Stronger selection then Hygro.
And there is also pheomycine... never used this before.
we used the neo selection pretty successfully.
V
V
Yes, in lots of general articles, I see that neo is frequently used. However, my PI isn't too keen on using Neo unless absolutely necessary since he said it takes a long long time to select...?
Thanks to both of you for your help
Dave
Thats true, its takes time to select neomycin.
U could try zeocin.