Team:UNIPV-Pavia/Methods Materials/Electrophoresis

From 2009.igem.org

(Difference between revisions)

Latest revision as of 10:30, 21 October 2009

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!!!)

INGREDIENTS:

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:

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-	<img width="100%" height="180px" src="https://static.igem.org/mediawiki/2009/c/c2/UNIPV_title_MandM.jpg" ></img>
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       <font face="Pristina" size="50" ><b>Protocols</b></font>
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+    <td width="50px"></td>
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 </table>
+<br>
+=='''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>
-Under Construction...
+*NOTE: when not specified, the marker has the following pattern:
+{|align="center"
+|[[Image:pv_ladder.jpg]]
+|}