Team:UNIPV-Pavia/Methods Materials

From 2009.igem.org

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<h1>Protocols</h1>
<h1>Protocols</h1>
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Lb"> LB medium preparation </a>
 
*[[Team:UNIPV-Pavia/Methods_Materials/Lb|Lb medium]]
*[[Team:UNIPV-Pavia/Methods_Materials/Lb|Lb medium]]
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*[[Team:UNIPV-Pavia/Protocols/Transformation|Transformation]]
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*[[Team:UNIPV-Pavia/Methods_Materials/Transformation|Transformation]]
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*[[Team:UNIPV-Pavia/Protocols/PlasmidExtraction|Plasmid extraction]]
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*[[Team:UNIPV-Pavia/Methods_Materials/XGal|X-Gal staining protocol for beta galactosidase (blue/white screening)]]
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*[[Team:UNIPV-Pavia/Protocols/Digestion|BioBrick digestion with restriction enzymes]]
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*[[Team:UNIPV-Pavia/Methods_Materials/Glycerol stock|Glycerol stock]]
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*[[Team:UNIPV-Pavia/Protocols/GelExtraction|DNA gel extraction]]
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*[[Team:UNIPV-Pavia/Methods_Materials/PlasmidExtraction|Plasmid extraction]]
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*[[Team:UNIPV-Pavia/Protocols/Precipitation|DNA precipitation with sodium acetate]]
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*[[Team:UNIPV-Pavia/Methods_Materials/Digestion|BioBrick digestion with restriction enzymes]]
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*[[Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase|Antarctic Phosphatase]]
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*[[Team:UNIPV-Pavia/Methods_Materials/GelExtraction|DNA gel extraction]]
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*[[Team:UNIPV-Pavia/Protocols/Ligation|Ligation]]
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*[[Team:UNIPV-Pavia/Methods_Materials/Ligation|Ligation]]
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*[[Team:UNIPV-Pavia/Protocols/Pcr|PCR]]
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*[[Team:UNIPV-Pavia/Methods_Materials/Pcr|PCR]]
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'''Gycerol stock'''
 
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*Mix 750 ul from the 5 ml sample of overnight incubated bacteria with 250 ul of 80% glycerol
 
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*Leave at -20°C for one day
 
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*Move to -80°C the day after
 
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'''X-Gal Staining Protocol for beta Galactosidase (blue/white screening)'''
 
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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.
 
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*Mix 20 ul X-Gal 20 mg/ml and 80 ul SOC and spread on LB+amp plates
 
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*Remember to use dark conditions
 
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*Incubate with 200 ul bacteria
 
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*Result: blue colonies express LacZ, while white colonies don't.
 
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Revision as of 14:47, 22 June 2009

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