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From 2010.igem.org

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Preparation of competent DH5α Escherichia coli cells

Autoclaving bottles & solutions for next day:

• Two 1000 ml empty bottles
• 500 ml 50 mM CaCl2 for 1000 ml cell culture
• 16 ml 50 mM CaCl2 + 15 % Glycerol

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Set a 5 ml LB ON culture of DH5α in 37°C, 250 rpm.