Inclusion bodies preparation
Sometimes you may want to try to purify your protein from  inclusion bodies for alternative purification procedures.

1. Grow 40ml of induced BL21 under best conditions for inclusion bodies (usually 37oc overnight in LB or 2xYT).
2. Pellet cells 10'  4,000rpm 4ºC in two 40ml tubes
3. Re-suspend in 1.2ml  lysis buffer leave 30min on ice
4. Sonicate 20'' x 3 (check the procedure)
5. Spin at 4ºC max speed 15' take sup into new tube (on ice)
6. Re-suspend in lysis buffer 0.5ml and sonicate 10''x 3 times
7. Spin max speed 15' transfer sup to new tube and re-suspend pellet in 0.1ml lysis buffer
8. Run samples from all lysates+pellets
9. Use the pellets for refolding experiments.

Lysis buffer
Tris pH 7.5 20mM
KCl  20mM
DTT   1mM
NP40  0.2%