Team:UNIPV-Pavia/Methods Materials/Electrophoresis

From 2009.igem.org

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== '''Electrophoresis''' ==  
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=='''Electrophoresis'''==
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''(estimated time: 2 hours)''
<br>
<br>
<br>
<br>
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<table width='80%'><tr><td>
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*Prepare agarose gel in 1x TBE buffer
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*Add ethidium bromide (using gloves and face mask for your safety):
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**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)
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*Cast the gel, insert the well-forming comb and let it polymerize
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*Add the loading buffer to each sample
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*Load the samples and 8 µl of marker
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*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
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*Take a picture of the gel, if needed (not when bands have to be cut!!!)
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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%'>
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'''TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)'''
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*'''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%'>
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'''Electrophoresis apparatus'''<hr color='white' width='70%'>
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'''Transilluminator'''</font>
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</td></tr></table></div><br><br><br><br>
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</td></tr></table>
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*NOTE: when not specified, the marker has the following pattern:
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{|align="center"
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|[[Image:pv_ladder.jpg]]
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|}

Revision as of 14:46, 14 October 2009

EthanolPVanimation.gif



Protocols


Electrophoresis

(estimated time: 2 hours)

  • 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!!!)










DNA samples
Ethidium bromide
Loading buffer 10x Blue Juice, Invitrogen

TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)

  • 54 gr Tris
  • 27.5 gr Borate
20 ml EDTA 0.5 M (pH 8)
Marker (1 kb DNA Ladder, Promega)
Face mask and gloves
Electrophoresis apparatus

Transilluminator





  • NOTE: when not specified, the marker has the following pattern:
Pv ladder.jpg
Retrieved from "http://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Electrophoresis"