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From 2012.igem.org

I. Inoculate Wild Type (WT) B. subtilis culture from Altman Lab in Nutrient Broth (Lim)

• Add ~12.5 mL of Nutrient broth to sterile 125 mL E-flask, re-cover with tin foil
• Inoculate 1 colony from pre-existing plate culture with inoculating loop to flask (11:05 AM)
• Grow at 37 ^oC with shaking overnight