From 2007.igem.org

Overexpression

1. Add 50 ml media (LB or minimal media) and 50 ul antibiotic (Carbenicillin) into flask.

2. Innoculate cells and incubate in 37°C shaker overnight.

3. Use UV/Vis spectroscopy to measure OD value.

4. Add enough culture into new flask to get a final OD of 0.5 and final volume of 50 ml by adding new media.

5. Add 1µM Isopropyl β-D-thiogalactopyranoside and incubate flask in 37°C shaker

6. After 24 hours, harvest the cells and store the cell pellet at - 80ْC