Yeast Crude Protein Extraction
Grow cells to mid-exponential phase in liquid medium.
Normalize cells to an OD600 of 2-3 in 1 mL of H2O.
Centrifuge 1 mL of H2O containing your cells at 9000 RPM for two minutes.
Resuspend pelleted cells in 100 uL 0.2 M NaOH and incubate on ice for 15 minutes.
Centrifuge cells at 9000 RPM for two minutes.
Resuspend the pellet in 100 uL of Yeast Protein Extraction Sample Buffer.
Add 0.4 uL 2-mercaptoethanol to the mixture.
Boil cells in sample buffer for 5 minutes at 95 C.
Can store at -80 C or load supernatant into a gel.
Vortex briefly then centrifuge at 10,000 RPM for 3 minutes.
