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
EDTA 1mM
DTT 1mM
NP40 0.2%