Dear Marthe,

Hygromycin is a tight selection as long as you remember the pH dependence

www.hygromycin.net/protocols.htm

We adjust our media to pH 7.5 AFTER AUTOCLAVING and select with 35ug/ml
hygromycin.

If your experiment allows, I suggest cotransforming the G418 and
hygromycin vectors.  The combined selection works better than either
selection alone.

best of luck,

Harry MacWilliams

----------------


On Thu, 18 Nov 2010, Ludtmann, MHR wrote:

> Dear all,
>
> I would like to overexpress a protein in a dicty cell line containing
> G418 resistance. Does anybody know of a suitable vector which is
> compatible with G418 resistance? I am aware of the pDE109 vector but I
> am worried about the reports of spontaneously resistant colonies.
>
> Any suggestions would be greatly appreciated.
>
> Thank you,
>
> Marthe
>
>
>
>
>
> Royal Holloway University of London
>
> Bourne building
>
> Egham
>
> TW20 0EX
>
>

----------------------------

Prof. Harry K. MacWilliams
Biologiezentrum der Ludwig-Maximilians-Universität
Grosshadenerstrasse 2
82152 Planegg-Martinsried
GERMANY

tel (office) +89 2180 74 288
tel (lab)    +89 2180 74 289
fax.         +89 2180 74 219

callers from North America please dial:
tel (office) 0114989 2180 74 288
tel (lab)    0114989 2180 74 289
fax          0114989 2180 74 219