Team:Macquarie Australia/WetLab/Protocols/Electrophoresis

From 2014.igem.org

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<h3>Agarose Gel Electrophoresis</h3>
<h3>Agarose Gel Electrophoresis</h3>
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<p>
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<h4> Preparing the Gel <h4>
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<h4> Preparing the Gel <h4>
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<li>Mix 1g of agarose powder with 100ml of 1x TAE buffer and heat for 1minute or until all agarose is dissolved.</li>
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Mix 1g of agarose powder with 100ml of 1x TAE buffer and heat for 1minute or until all agarose is dissolved.
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<li>Wait until it has cooled (not set), and add 1ul of GelRed into the mixture. </li>
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</li>
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<li>Pour the solution into a cast with an appropriate comb.</li>
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<li>
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<li>Leave to set.</li>
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Wait until it has cooled (not set), and add 1ul of GelRed into the mixture. </li>
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</ul>
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<li>
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Pour the solution into a cast with an appropriate comb.</li>
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<li>
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Leave to set</li>
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<h4> Running the Gel </h4>
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<h4> Running the Gel </h4>
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<ul style="list-style-type: decimal;">
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<li>Mix 1ul of 1kbp DNA ladder with 6ul of loading dye (bromophenol blue) and 4ul of 1x TAE buffer (total 6ul) and load onto first well </li>
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<li>Mix 1ul of 1kbp DNA ladder with 6ul of loading dye (bromophenol blue) and 4ul of 1x TAE buffer (total 6ul) and load onto first well </li>
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<li>Mix 5ul of PCR products with 1ul of loading dye and load onto wells.</li>
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<li>Mix 5ul of PCR products with 1ul of loading dye and load onto wells.</li>
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<li>Run gel at 90V for 45minutes approximately</li>
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<li>Run gel at 90V for 45minutes approximately</li>
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<li>Photograph gels under UV light</li>
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<li>Photograph gels under UV light</li>
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</ul>
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Revision as of 04:46, 16 October 2014

Protocols


Agarose Gel Electrophoresis

Preparing the Gel

  • Mix 1g of agarose powder with 100ml of 1x TAE buffer and heat for 1minute or until all agarose is dissolved.
  • Wait until it has cooled (not set), and add 1ul of GelRed into the mixture.
  • Pour the solution into a cast with an appropriate comb.
  • Leave to set.

Running the Gel

  • Mix 1ul of 1kbp DNA ladder with 6ul of loading dye (bromophenol blue) and 4ul of 1x TAE buffer (total 6ul) and load onto first well
  • Mix 5ul of PCR products with 1ul of loading dye and load onto wells.
  • Run gel at 90V for 45minutes approximately
  • Photograph gels under UV light

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