Equipment:

- Incubation shaker

- Photometer

Chemicals & consumables:

- E. coli BL21 DE3

- dYT medium or LB medium

- IPTG

- 100 ml and 3 l flasks

- Ice

Procedure:

Inoculation of 50 mL LB medium in a 100 mL flask with E. coli BL21 DE3 containing the plasmid with the sequence for the respective part to be expressed.

Incubation at 180 repulsion per minute (rpm) at 30°C to an OD₆₀₀= 4

Transferation of the starter culture into 1 L LB medium in a 3 L flask resulting in an OD₆₀₀= 0.2

Incubation to an OD₆₀₀= 0.6 at 180 rpm and 30°C.

Incubation for 15 minutes on ice.

Induction of the protein expression with 20 mL of IPTG (stock concentration 1M).

Incubation of the cell suspension overnight at 180 rpm at 30°C.