Team:UNIPV-Pavia/Methods Materials/Electrophoresis
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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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+ | =='''Electrophoresis'''== | ||
+ | ''(estimated time: 2 hours)'' | ||
+ | <br> | ||
+ | <br> | ||
+ | <table width='80%'><tr><td> | ||
+ | *Prepare agarose gel in 1x TBE buffer | ||
+ | *Add ethidium bromide (using gloves and face mask for your safety): | ||
+ | **1 µl in the small size agarose gel (70 ml) | ||
+ | **2 µl in the middle size agarose gel (150 ml) | ||
+ | **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 | ||
+ | *Use UV-light to look at the bands | ||
+ | *Take a picture of the gel, if needed (not when bands have to be cut!!!) | ||
+ | <br><br><br><br><br><br><br><br> | ||
+ | <br></td><td align='center' valign='top' width='300px'> | ||
+ | |||
+ | <table border='1'> | ||
+ | <tr bgcolor='yellow'><td>INGREDIENTS: | ||
+ | </td></tr> | ||
+ | <tr><td> | ||
+ | <font class='label'> | ||
+ | '''DNA samples'''<hr color='white' width='70%'> | ||
+ | '''Ethidium bromide'''<hr color='white' width='70%'> | ||
+ | '''Loading buffer 10x Blue Juice, Invitrogen'''<hr color='white' width='70%'> | ||
+ | '''TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)''' | ||
+ | *'''54 gr Tris''' | ||
+ | *'''27.5 gr Borate'''<hr color='white' width='70%'> | ||
+ | '''20 ml EDTA 0.5 M (pH 8)'''<hr color='white' width='70%'> | ||
+ | '''Marker (1 kb DNA Ladder, Promega)'''<hr color='white' width='70%'> | ||
+ | '''Face mask and gloves'''<hr color='white' width='70%'> | ||
+ | '''Electrophoresis apparatus'''<hr color='white' width='70%'> | ||
+ | '''Transilluminator'''</font> | ||
+ | </td></tr></table><br><br><br><br> | ||
+ | </td></tr></table> | ||
- | + | *NOTE: when not specified, the marker has the following pattern: | |
+ | {|align="center" | ||
+ | |[[Image:pv_ladder.jpg]] | ||
+ | |} |
Latest revision as of 10:30, 21 October 2009
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Electrophoresis
(estimated time: 2 hours)
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- NOTE: when not specified, the marker has the following pattern:
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