Yo (how's that for an all-inclusive greeting?),
Last week I posted a question with regard to our inability to get 5FOA to
work on a defined minimal medium (as opposed to the standard YNB-based
medium). Here is a summary of the responses.
Vince Schulz- The trick is to leave out the buffering agents in synthetic
media (ie succinic acid and NaOH).
Tom Chappell- ....in my hands, both EMM and dropout 5FOA
kill ura+ cells.
(I was using PM, so there may be a difference between the two.)
Bill Clinton- subscribe yeast (How did that get in here?)
Other responses from Tom Chappell and from Karen Hahnenberger addressed my
direct concern that I could not turn on the nmt (no message on thiamine)
promoter on a YNB-based medium. They both indicated that YNB contains
insufficient thiamine for complete repression of nmt transcription.
Thanks to all who responded.
"I'd rather be fission."