Yeast Protein Expression

3/28/01

1. Grow yeast to saturation O/N to O/ 2 nights

2. Dilute 1:10 into either growth medium (+glucose for bait) or induction medium (+GR for library plasmid)

3. Grow to OD=0.5 (3 hours for glucose)

4. spin 10’ 2500RPM tc fuge

5. add 50uL 2X sample buffer

6. snap freeze on dry ice

7. boil 10’

8. sonicate 10 seconds

9. spin 10’ 14K

10. Run on 10% gel and transfer

11. Blot for protein expression – library anti-HA (mouse)

bait anti-LexA (mouse)