Protein Expression Using NiCo21(DE3) (C2529)
- Transform expression plasmid into NiCo21(DE3). Plate on antibiotic selection plates and incubate overnight at 37°C.
- Resuspend a single colony in liquid culture with antibiotic to produce a starter culture. Inoculate starter culture at a 1:100 dilution into expression media containing antibiotic.
- Incubate at 37°C with shaking until OD600 reaches 0.4–0.8.
- For most vector systems, induce expression with 40 or 400 µM IPTG for 3 hours at 37°C, 5 hours at 30°C or overnight at 16°C or 23°C.