Problems with beta-gal assay - need help.

Seng-Lai Tan sltan at u.washington.edu
Mon Aug 5 19:04:56 EST 1996


we are having the same problem with b-gal assays of the yeast 2-hybrid
system.  things that turned dark blue now are longer blue.  we think, as
well as stanley fields himself, that the lacZ reporter is probably kicked
out from the yeast strains because there isn't any selection for it
whereas the his3 reporter gene is constantly under selection on minus
histidine medium.  so are you applying selection for your lacZ reporter?

On 5 Aug 1996, mordechai choder wrote:

> We are routinely performing beta-gal assays of yeast. We have used glass
> beads (Biospec) extraction (no chloroform no SDS) and obtained very
> reproducible results... until recently when everything started to fail.
> Results are not results, duplicates are not duplicate and in 50-90% of the
> time color simply does not develop.  We changed everything possible with no
> improvement.  Seek for advice and suggestions. Thanks.
> Mordechai Choder.
>
>
>
>


Thomas Seng-Lai Tan               ...................................
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