Dear Dicty fans,

Hygromycin toxicity is very strongly dependent on pH (see
www.hygromycin.net/protocols.htm).  At pH 7,5 one can select Dicty
multicopy transformants effectively with 30-40 ug/ml hygromycin. Note that
autoclaving lowers the pH, so one either has to filter-sterilize
or re-adjust the pH of the sterilized medium.  Since hygro is expensive,
attention to pH can also result in considerable savings.

best wishes

Harry

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On Fri, 27 Feb 2009, Vanessa Mcmains wrote:

> Dicty community,
>
> I am using hygromycin as a selectable marker for overexpression of a
> gene. I do not see high expression using 50ug/ul hygromycin for
> selection. What is the highest concentration that anyone has used in
> Dicty to successfully select for overexpressing cells using
> hygromycin?
>
> Thanks,
> Vanessa
>

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

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