Yes, expression of proteins at 18°C and 25°C has been validated using the Dual Temperature protocol as follows. Day 1: Inoculate LB + antibiotic with an expression colony and incubate overnight at 30°C, 300 rpm. Day 2: Inoculate pre-warmed (30°C) MagicMedia + antibiotic at 1:40 with overnight seed culture. Incubate culture at 30°C, 300 rpm for 6-7 hours or until OD600 reaches >6. Transfer culture to either 18°C or 25°C, 300 rpm and incubate for 24-36 hours. Day 3: Harvest and analyze results.
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