Hygromycin B and Cells

[Glenn Jenkins]

In article <51os0v$1de6 at news.rhrz.uni-bonn.de>,
dramiga at snphysio2.wilhelm.uni-bonn.de (Labor Wernig) wrote:

> Hello Netters !
> 
> Does anyone know the usual working concentration for hygromycin B ?
> I transformed E.coli with a plasmid containing a hygromycin B resistance
gene, but unfortunately also the untransformed bacteria survived when they
were grown in medium containing hygromycin B.
> I would also like to work with mouse myoblasts. Do I have to use
different working concentrations for mammalian cells and E.coli ?
> 
> Any help would be much appreciated
> 
> Joe Otim Dramiga
> 
> E-Mail: dramiga at snphysio2.wilhelm.uni-bonn.de

I have never heard of using hygro in bacteria. Although I have no idea
what plasmid you are using. Are you sure this gene has a bacteral promoter
in front of it? Typically Hygro is used for mammalian cells at a
concentration 
around 200 ug/ml. You should titrate this amount on non-transfected cells
first to be sure.

Glenn