Team:UNIPV-Pavia/Methods Materials

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Gycerol stock

• Mix 750 ul from the 5 ml sample of overnight incubated bacteria with 250 ul of 80% glycerol
• Leave at -20°C for one day
• Move to -80°C the day after

X-Gal Staining Protocol for beta Galactosidase (blue/white screening) The principle is that X-Gal (5-bromo-4-chloro-3-indolyl-b-D-galactopyranoside) turns blue when reacts with beta-galactosidase.

• Mix 20 ul X-Gal 20 mg/ml and 80 ul SOC and spread on LB+amp plates
• Remember to use dark conditions
• Incubate with 200 ul bacteria
• Result: blue colonies express LacZ, while white colonies don't.