Lissa1:Yeast Protein Extracts

Solution 1:
 * 1.85 M NaOH
 * 7.3% B-mercaptoethanol
 * Must make this fresh!

Solution 2:
 * 50% TCA (store 4 degrees)

Solution 3:
 * Acetone (store -20 degrees)

Sample buffer:
 * 1.25 mL 4x Laemmli protein loading buffer
 * .25 mL 1 M DTT
 * .15 mL Tris base
 * 3.35 mL water
 * Either make fresh or store.

Steps:
 * 1) Pellet 1.5 mL of cell culture having OD 1.5.
 * 2) Remove supernatant.
 * 3) Freeze in liquid nitrogen and store at -80 degrees.
 * 4) Add 150 uL of Sol. 1 to each tube, vortex to resuspend.
 * 5) Incubate 10 min on ice.
 * 6) Add 150 uL of Sol. 2 and invert to mix.
 * 7) Incubate 10 min on ice.
 * 8) Centrifuge 2 min at 4 degrees, remove supernatant.
 * 9) Rinse pellet with 1 mL of Sol. 3.
 * 10) Centrifuge 2 min at 4 degrees, remove supernatant.
 * 11) Dry tubes upside down under a tube tray on a paper towel for 3 min.
 * 12) Add 50 uL sample buffer.
 * 13) Break up pellet with tip. Vortex.
 * 14) Boil in sand-boiler for 5 min, vortex again, load around 20 uL on sumo gel.