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Team:UNIPV-Pavia/Methods Materials/XGal
From 2009.igem.org
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<font face="Pristina" size="50" ><b>Protocols</b></font> | <font face="Pristina" size="50" ><b>Protocols</b></font> | ||
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*The principle is that X-Gal (5-bromo-4-chloro-3-indolyl-b-D-galactopyranoside) turns blue when reacts with beta-galactosidase. | *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 (X-Gal and DMF are toxic, use face-mask for your safety!!! X-Gal is light-sensitive, remember to keep it in the dark, when possible) | *Mix 20 ul X-Gal 20 mg/ml and 80 ul SOC and spread on LB+amp plates (X-Gal and DMF are toxic, use face-mask for your safety!!! X-Gal is light-sensitive, remember to keep it in the dark, when possible) | ||
*Incubate with 200 ul bacteria | *Incubate with 200 ul bacteria | ||
*Result: blue colonies express LacZ, while white colonies don't. | *Result: blue colonies express LacZ, while white colonies don't. | ||
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| + | <tr bgcolor='yellow'><td>INGREDIENTS: | ||
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| + | '''40 mg/ml X-gal in DMF'''<hr color='white' width='70%'> | ||
| + | '''SOC medium'''<hr color='white' width='70%'> | ||
| + | '''Pre-warmed LB agar plate'''<hr color='white' width='70%'> | ||
| + | '''Face mask'''</font> | ||
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Latest revision as of 10:27, 21 October 2009
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X-Gal staining protocol for beta galactosidase (blue/white screening)
(Estimated time: 20 min)
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