pONE-10K

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	For protein expression in bacteria, E. coli BL21 (DE3) cells were transformed with the appropriate plasmids and plated on antibiotic-containing Luria-Bertani broth (LB; plus 50 μg/mL kanamycin or 100 μg/mL ampicillin). A selected colony was inoculated in 20 mL LB medium and incubated overnight at 37°C, 220 rpm in the presence of an antibiotic.

Protocol tips
For protein expression in bacteria, E. coli BL21 (DE3) cells were transformed with the appropriate plasmids and plated on antibiotic-containing Luria-Bertani broth (LB; plus 50 μg/mL kanamycin or 100 μg/mL ampicillin). A selected colony was inoculated in 20 mL LB medium and incubated overnight at 37°C, 220 rpm in the presence of an antibiotic.

Publication protocol

For protein expression in bacteria, E. coli BL21 (DE3) cells were transformed with the appropriate plasmids and plated on antibiotic-containing Luria-Bertani broth (LB; plus 50 μg/mL kanamycin or 100 μg/mL ampicillin). A selected colony was inoculated in 20 mL LB medium and incubated overnight at 37°C, 220 rpm in the presence of an antibiotic. Next day, 1 mL or 10 mL of the starter culture was used to inoculate 100 mL or 1 L of LB broth respectively and cells were grown at 37°C, 220 rpm in the presence of an antibiotic. Protein expression was induced by adding 1 mM IPTG to the cell cultures, upon reaching an OD 600 density of ~0.8–0.9. Before induction, cultures were equilibrated to induction temperature 30°C for 0.5 hours. Protein expression was continued for 4 hours at 30°C, 220 rpm.

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