2 replies to this topic

[Thomson]

Hi, 

 

I've been given a plasmid, and the vector map shown the antibiotic resistance gene marked 'Neo/KanR'.

Just for confirmation does it mean the gene translation products can resistant to both Neomycin and Kanamycin. 

I only need to use either one antibiotic for selection purpose right?

 

Also, anyone know the suitable final concentration of Kanamycin to be used for selection?

Some told me 25 ug/ul, some told me 50 ug/ul. I was told too high might cause the bacteria to not able to grow at all.

 

Thanks

[bob1]

You only need to use one - generally Kan is used as it is cheaper. The Neo gene also confers antibiotic resistance for eukaryotic cells, so that you can do selection of transfected cells.

 

I would use Kan at 50 ug/ml (not ug/ul as you have above - that would make a good stock concentration though for a 1:1000 dilution to working), but if you have a low copy number plasmid then you should use at 25 ug/ml.

[Thomson]

Thanks. Sorry for the typo. I'd try with 50ug/ml final concentration.