Team:UiOslo Norway/Experiments/Protein Expression

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Protein Expression:

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• Inoculate 5 ml preculture and incubate at 37 °C overnight shaking at 220 rpm

• Measure OD₆₀₀ and calculate the amount needed to inoculate an expression culture with and OD₆₀₀ of 0.3

• Inoculate the expression culture and incubate at 37 °C for 1 hour shaking at 220 rpm

• Determine OD₆₀₀

• Recommended is an OD₆₀₀ between 0.4-1.0 to induce protein expression

• Take a 1 ml sample in an uninduced stage

• Induce protein expression by adding IPTG to a final concentration of 1 mM

• Incubate the culture for 3 hours at 37 °C shaking at 220 rpm

• Harvest cells by centrifugation at 8,000 x g for 5 minutes