thiamine question

Colin Rasmussen colin at pombe.usask.ca
Fri Feb 4 12:31:57 EST 2000



"Long, Kimberly" wrote:

> Hi,
>
> I am a new graduate student working with S. pombe.  My favorite gene is
> under the control of the nmt1 promoter and I am doing a repression
> experiment over a long time course.  Can anyone tell me how long thiamine
> lasts in media at 30 C?  Do I need to replenish the thiamine after a certain
> amount of time?

We use 1 uM and do experiments that last over several days.  We've been able to
grow cells from 10E5 cells per ml to saturation density without noticing any
loss of repression.  If there had been a loss we'd have known as we were
growing cells which express lethal proteins.

Colin Rasmussen
U of Saskatchewan

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