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about antibiotic in plasmid transformation - which is best? (Mar/17/2006 )

I have a plsmid with amp, kana anti-sites and a GFP marker, but I want to know I can select which antibiotics, either amp or kana, or both amp and kana in transform process.

Many Thanks,

Max smile.gif

-chiangmax-

You can select with either one, or even with both together (in this case I would maybe lower the concentration of both a bit, but I don't see why you would use both as one will do).

-vairus-

QUOTE (chiangmax @ Mar 18 2006, 12:23 AM)
I have a plsmid with amp, kana anti-sites and a GFP marker, but I want to know I can select which antibiotics, either amp or kana, or both amp and kana in transform process.

Many Thanks,

Max smile.gif


Thanks for your advice

max

-chiangmax-

If you have to choose one, Kan is better because unlike Amp it"s not degraded, so you won"t see the micro satelittes around the main colony that usually appear when using amp. You can also replace amp by carbenicillin (I am not 100% sure of spelling) which is a wee bit more stable, but allow growth of AmpR strains.

-Canalon-

The Kanamycin gene has (also) a bacterial promoter? I mean, it is not (only) for expression in eukaryotes for neo selection?

-il0postino-

Appreciate to your answers which must be helpful to my recent study. If positive results appear I will tell all of you.

Many thanks,

Max smile.gif

-chiangmax-

QUOTE (il0postino @ Mar 25 2006, 02:13 PM)
The Kanamycin gene has (also) a bacterial promoter? I mean, it is not (only) for expression in eukaryotes for neo selection?


Probably depends on the vector. But it can be usefull in bacteria to have a selection marker other than amp, so it makes sense, and many commercial vectors have both.

-Canalon-