http://2009.igem.org/wiki/index.php?title=Team:Waterloo/Notebook/Protocols/Gel_Electrophoresis&feed=atom&action=historyTeam:Waterloo/Notebook/Protocols/Gel Electrophoresis - Revision history2024-03-29T04:36:32ZRevision history for this page on the wikiMediaWiki 1.16.5http://2009.igem.org/wiki/index.php?title=Team:Waterloo/Notebook/Protocols/Gel_Electrophoresis&diff=166759&oldid=prevTimlin737: New page: {{Team:Waterloo/NavBar}} <b><u><big>Gel Electrophoresis</big></u></b><br> <b>Materials </b><br> Agarose <br> 1X TAE buffer <br> Gel Red<br> 1:10 diluted 1kb DNA ladder<br> Loading Dye<...2009-10-22T02:54:41Z<p>New page: {{Team:Waterloo/NavBar}} <b><u><big>Gel Electrophoresis</big></u></b><br> <b>Materials </b><br> Agarose <br> 1X TAE buffer <br> Gel Red<br> 1:10 diluted 1kb DNA ladder<br> Loading Dye<...</p>
<p><b>New page</b></p><div>{{Team:Waterloo/NavBar}}<br />
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<b><u><big>Gel Electrophoresis</big></u></b><br> <br />
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<b>Materials </b><br><br />
Agarose <br><br />
1X TAE buffer <br><br />
Gel Red<br><br />
1:10 diluted 1kb DNA ladder<br><br />
Loading Dye<br><br />
Gel Rig<br><br />
<br />
<b>Instructions for 0.8% Agarose 100 mL gel</b><br><br />
<u>Prepare the Gel </u><br><br />
1. Measure 0.8 g of agarose on weigh paper. <br><br />
2. Measure 100mL of TAE buffer using a graduated cylinder into a flask. <br><br />
3. Add agarose to flask. Swirl to dissolve as much as possible. <br><br />
4. Microwave for 20 seconds, swirl. Repeat until agarose is dissolved. Allow argrose to cool down to room temperature.<br><br />
5. Add 2 µL of Gel Red into the gel.<br><br />
6. Cover the ends of the gel tray with masking tape to seal them. <br><br />
7. Pour the gel into the tray. <br><br />
8. Drop the combs into the slots to form the wells. <br><br />
9. Allow the gel to set. <br><br />
10. Remove the combs and rinse with distilled water. <br><br />
<br />
<u>Preparing samples </u><br><br />
1. Pipette 10µL each digested sample into a labeled microfuge tube (for gel extraction use 20 µL). <br><br />
2. Add 2µL of loading dye to each sample (for gel extraction use 4 µL) <br><br />
3. Find the diluted 1kb DNA ladder. <br><br />
4. Organise the tubes in loading order.<br><br />
5. Record loading order <br><br />
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<u>Running the Gel </u><br><br />
1. Place the tray in the rig with the wells closest to the far side. <br><br />
2. Add TAE buffer until the surface of the gel is just covered. <br><br />
3. Pipette 10 µL of the digestion mixture and diluted ladders into the appropriate wells.<br><br />
4. Put the cover on. Black at the back and red at the head. <br><br />
5. Connect the wires to an available power supply. <br><br />
6. Set the power supply to 120V and start it. <br><br />
7. Check that bubbles are forming near the electrodes. <br><br />
8. Wait about 45 minutes. The dye front should be 3/4 of the way down the gel.<br></div>Timlin737