Team:UNIPV-Pavia/Methods Materials/XGal

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(New page: {{UNIPV-Pavia/Protocols}} <br> == '''X-Gal staining protocol for beta galactosidase (blue/white screening)''' == ''(Estimated time: 20 min)'' <br> <br> '''Materials needed:''' *'''40 mg/m...)
(→X-Gal staining protocol for beta galactosidase (blue/white screening))
 
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== '''X-Gal staining protocol for beta galactosidase (blue/white screening)''' ==  
== '''X-Gal staining protocol for beta galactosidase (blue/white screening)''' ==  
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'''Materials needed:'''
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*'''40 mg/ml X-gal in DMF'''
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*'''SOC medium'''
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*'''pre-warmed LB agar plate'''
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*'''face mask'''
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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%'>
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'''SOC medium'''<hr color='white' width='70%'>
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'''Pre-warmed LB agar plate'''<hr color='white' width='70%'>
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Latest revision as of 10:27, 21 October 2009

EthanolPVanimation.gif



Protocols


X-Gal staining protocol for beta galactosidase (blue/white screening)

(Estimated time: 20 min)

  • 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)
  • Incubate with 200 ul bacteria
  • Result: blue colonies express LacZ, while white colonies don't.










INGREDIENTS:

40 mg/ml X-gal in DMF
SOC medium
Pre-warmed LB agar plate

Face mask





Retrieved from "http://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/XGal"