Induction test of a recombinant protein
1. 2 ml ON culture of BL21 transformed with the expression construct in LB ampi.
2. Inoculate 20ml LB+ampi with the ON culture and incubate until OD600 = 0.5/0.6.
3. Separate the 20ml into 2X10ml and add 400μM IPTG to only one of the cultures. Incubate 2h30 at 37 or 3h at 25°C (or ON at 16°C.
4. Spin down 1ml of each culture and ressuspend the pellet in 200ul of cracking buffer.
5. Heat at 95°C during 4min and SDS PAGE.
Cracking buffer 3X: Glycerol 30%
BME 2%
SDS 3%
Tris HCl pH 6.6 150mM
Bromophenol blue 0,001%
2. Inoculate 20ml LB+ampi with the ON culture and incubate until OD600 = 0.5/0.6.
3. Separate the 20ml into 2X10ml and add 400μM IPTG to only one of the cultures. Incubate 2h30 at 37 or 3h at 25°C (or ON at 16°C.
4. Spin down 1ml of each culture and ressuspend the pellet in 200ul of cracking buffer.
5. Heat at 95°C during 4min and SDS PAGE.
Cracking buffer 3X: Glycerol 30%
BME 2%
SDS 3%
Tris HCl pH 6.6 150mM
Bromophenol blue 0,001%