alternative to raffinose?

Sailesh Surapureddi Saisu at mcb.liu.se
Mon Jun 2 03:51:53 EST 1997



Hi Hendri

		I had the same problem and Bob (the guy who made those
yeast vectors) sugggested the following.. I am pasting as it is..
if you need further clarifications, pls feel free to mail me directly.

So here are your options:

1)   Grow on  SC-uracil-glucose to mid-log, spin and wash cells, then add
YEP-4%g galactose and induce
for greater than 6 hours.  This should be ample time to overcome glucose
repression and induction
of the GST fusion promoter.

2)   Grow in SC-uracil-glucose (2%) to establish a starter culture.  Use
this to start a culture in
YEP-Raffinose (2%) or YEP-glycerol/ethanol (2%/2%).  At mid-log, simply add
galactose to the
culture to make the final concetration 4%.  You will begin seeing induction
within minutes, but for
maximum yield (assuming no toxicity), induce for greater than 6 hours.


Good luck

Sailesh.




Sailesh Surapureddi Ph.D		Communicators
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