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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 | ||
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| + | '''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. | ||
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Revision as of 14:27, 22 June 2009
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Gycerol stock
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.
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