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| | + | {{UNIPV-Pavia/StyleCss}} |
| | <html> | | <html> |
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| | <td width="50%"></td> | | <td width="50%"></td> |
| | <td align="center"><br><bR> | | <td align="center"><br><bR> |
| - | <font face="Pristina" size="50" ><b>Protocols</b></font> | + | <font face="Pristina" size="50" ><b>Instruments</b></font> |
| | </td> | | </td> |
| | </html> | | </html> |
| | </tr> | | </tr> |
| | <tr> | | <tr> |
| - | <td> {{UNIPV-Pavia/Protocols}}</td> | + | <td> {{UNIPV-Pavia/Instruments}}</td> |
| | </tr> | | </tr> |
| | </table> | | </table> |
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| | <br> | | <br> |
| - | =='''Electrophoresis'''== | + | == ''Water bath'' == |
| - | ''(estimated time: 2 hours)''
| + | |
| | <br> | | <br> |
| | <br> | | <br> |
| - | <table width='80%'><tr><td>
| + | {| |
| - | *Prepare agarose gel in 1x TBE buffer
| + | |[[Image:pv_vasche.jpg|thumb|350px|left|Electrophoresis instrumentation]] |
| - | *Add ethidium bromide (using gloves and face mask for your safety):
| + | |
| - | **1 µl in the small size agarose gel (70 ml)
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| - | **2 µl in the middle size agarose gel (150 ml)
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| - | **4 µl in the big size agarose gel (250 ml)
| + | |
| - | *Cast the gel, insert the well-forming comb and let it polymerize
| + | |
| - | *Add the loading buffer to each sample
| + | |
| - | *Load the samples and 8 µl of marker
| + | |
| - | *Set to 70-100 volts and electrophorese for the required amount of time
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| - | *Use UV-light to look at the bands
| + | |
| - | *Take a picture of the gel, if needed (not when bands have to be cut!!!)
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| - | <br><br><br><br><br><br><br><br>
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| - | <br></td><td align='center' valign='bottom' width='300px'>
| + | |
| - | <div align='center'>
| + | |
| - | <table border='1'>
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| - | <tr><td>
| + | |
| - | <font class='label'>
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| - | '''DNA samples'''<hr color='white' width='70%'>
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| - | '''Ethidium bromide'''<hr color='white' width='70%'>
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| - | '''Loading buffer 10x Blue Juice, Invitrogen'''<hr color='white' width='70%'>
| + | |
| - | '''TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)'''
| + | |
| - | *'''54 gr Tris'''
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| - | *'''27.5 gr Borate'''<hr color='white' width='70%'>
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| - | '''20 ml EDTA 0.5 M (pH 8)'''<hr color='white' width='70%'>
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| - | '''Marker (1 kb DNA Ladder, Promega)'''<hr color='white' width='70%'>
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| - | '''Face mask and gloves'''<hr color='white' width='70%'>
| + | |
| - | '''Electrophoresis apparatus'''<hr color='white' width='70%'>
| + | |
| - | '''Transilluminator'''</font>
| + | |
| - | </td></tr></table></div><br><br><br><br>
| + | |
| - | </td></tr></table>
| + | |
| - | | + | |
| - | *NOTE: when not specified, the marker has the following pattern:
| + | |
| - | {|align="center" | + | |
| - | |[[Image:pv_ladder.jpg]] | + | |
| | |} | | |} |
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