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| | {{UNIPV-Pavia/Menu}} | | {{UNIPV-Pavia/Menu}} |
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| - | <h1>Protocols</h1> | + | <td align="center"><br><bR> |
| - | | + | <font face="Pristina" size="50" ><b>Protocols</b></font> |
| - | <a href="https://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Lb"> LB medium preparation </a> | + | |
| - | *[[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/Protocols/PlasmidExtraction|Plasmid extraction]]
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| - | *[[Team:UNIPV-Pavia/Protocols/Digestion|BioBrick digestion with restriction enzymes]]
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| - | *[[Team:UNIPV-Pavia/Protocols/GelExtraction|DNA gel extraction]]
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| - | *[[Team:UNIPV-Pavia/Protocols/Precipitation|DNA precipitation with sodium acetate]]
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| - | *[[Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase|Antarctic Phosphatase]]
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| - | *[[Team:UNIPV-Pavia/Protocols/Ligation|Ligation]]
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| - | *[[Team:UNIPV-Pavia/Protocols/Pcr|PCR]]
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| - | <br> | + | |
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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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| | </td> | | </td> |
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| | </tr> | | </tr> |
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| | </table> | | </table> |
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