Hello Wim,
what about you screen them after your transformation on SD -W -L -H +3AT
and then grid the upcoming colonies onto SD -W -L to get colonies big enough
for the filter assay?
Just an idea, but please let me know in case this helps,
good luck!
Daniel
Daniel Schlieper, Institute of Genetics, University of Cologne,
Zuelpicher Str. 67, D-50674 Cologne, Germany
Tel.: ++49 (0)221 470 4848 FAX: ++49 (0)221 470 5170
Email: dschliep at genetik.uni-koeln.de
>From: wvcrieki at lmb1.rug.ac.be>To: "bionet.molbio.yeast mail newsgroup" <bionet-news at dl.ac.uk>
>Subject: Two Hybrid Screening Problem
>X-Article-Number: bionet.molbio.yeast Msg # 2627
>>Hello ...,
>>In using CG-1945 for a two hybrid screening I'm having trouble in obtaining
>colonies big enough to lift (for the lysis in liquid nitrogen). The colonies
>start growing after 2-3 days but stop after 5 days. How can I overcome this ?
>e.g. changing X-gal assay, or using different plating techniques (I'm using SD
>plates lacking LEU,TRP & HIS, 5 mM 3-TA). All suggestions are welcome.
>>Thanks for your time
>Wim Van Criekinge
>E-mail:Wvcrieki at lmb1.rug.ac.be
