Mini Expression

We used this protocol to define the optimum conditions for expression of our constructs.

Reagents and Materials

• Overnight cultures of protein-producing bacteria
• LB broth
• IPTG (Isopropyl-Beta-D-Thiogalactoside)

Protocol

1. Make 2mL overnight cultures of your protein-producing bacteria
2. Subculture in 10mL of LB broth
3. When OD600 is between 0.3 and 0.5, induce the culture with IPTG
4. To determine optimum conditions for expression, use different time point of induction (e.g. 4h, 8h and 12h) and different concentration of IPTG (e.g., 0.1mM, 0.4mM and 0.8mM)