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E. coli W3110 cells harboring the expression vectors were grown with agitation in 1 l erlenmeyer flaks containing 100 ml of Luria-Bertani medium supplemented with glycerol (10 g/l) kanamycin (50 mg/l) at 30°C. Protein expression was induced when OD600 reached 0.5 units by adding 2 g/l L-arabinose and incubation continued for an additional 5 h period. Final OD600 was typically between 8-10 units. |
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| Protocol tips |
| E. coli W3110 cells harboring the expression vectors were grown with agitation in 1 l erlenmeyer flaks containing 100 ml of Luria-Bertani medium supplemented with glycerol (10 g/l) kanamycin (50 mg/l) at 30°C. Protein expression was induced when OD600 reached 0.5 units by adding 2 g/l L-arabinose and incubation continued for an additional 5 h period. Final OD600 was typically between 8-10 units. |
Publication protocol
E. coli W3110 cells harboring the expression vectors were grown with agitation in 1 l erlenmeyer flaks containing 100 ml of Luria-Bertani medium supplemented with glycerol (10 g/l) kanamycin (50 mg/l) at 30°C. Protein expression was induced when OD600 reached 0.5 units by adding 2 g/l L-arabinose and incubation continued for an additional 5 h period. Final OD600 was typically between 8-10 units. Cells were then harvested by centrifugation at 10,000g for 15 min at 4°C.
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