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Team:Macquarie Australia/WetLab/Protocols/Electrophoresis
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<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/CompetentCells">Competent Cells</a></li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/CompetentCells">Competent Cells</a></li> | ||
<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Electrophoresis">Electrophoresis</a></li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Electrophoresis">Electrophoresis</a></li> | ||
| - | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Ligation">Ligation</a></<li> | + | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Ligation">Composite Part <br/>Ligation</a></<li> |
<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Trypsin">Manual Trypsin <br/>In-Gel Digestion</a></<li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Trypsin">Manual Trypsin <br/>In-Gel Digestion</a></<li> | ||
<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/MassSpec">Mass Spectrometry</a></li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/MassSpec">Mass Spectrometry</a></li> | ||
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<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/PlasmidPreps">Plasmid Prep</a></li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/PlasmidPreps">Plasmid Prep</a></li> | ||
<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/SDSPAGE">SDS-PAGE</a></li> | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/SDSPAGE">SDS-PAGE</a></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Heat">Heat Shock</a></li> | ||
| + | <li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Operons">Induction of Operons</a></li> | ||
</ul> | </ul> | ||
</div> | </div> | ||
<div class="cont-out"> | <div class="cont-out"> | ||
| - | <h3>Electrophoresis</h3> | + | <h3>Agarose Gel Electrophoresis</h3><br/> |
| - | <p></p> | + | <h4> Preparing the Gel </h4> |
| + | <ul style="list-style-type: decimal;"> | ||
| + | <li>Mix 1g of agarose powder with 100ml of 1x TAE buffer and heat for 1minute or until all agarose is dissolved.</li> | ||
| + | <li>Wait until it has cooled (not set), and add 1ul of GelRed into the mixture. </li> | ||
| + | <li>Pour the solution into a cast with an appropriate comb.</li> | ||
| + | <li>Leave to set.</li> | ||
| + | </ul> | ||
| + | |||
| + | <h4> Running the Gel </h4> | ||
| + | <ul style="list-style-type: decimal;"> | ||
| + | <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> | ||
| + | <li>Mix 5ul of PCR products with 1ul of loading dye and load onto wells.</li> | ||
| + | <li>Run gel at 90V for 45minutes approximately</li> | ||
| + | <li>Photograph gels under UV light</li> | ||
| + | </ul> | ||
| + | |||
| + | <img src="https://static.igem.org/mediawiki/2014/b/b4/Agarose.png" /> | ||
| + | <p><b>Figure 1.</b> Bio-Rad agarose gel.</p> | ||
| + | |||
</div> | </div> | ||
</section> | </section> | ||
</body> | </body> | ||
</html> | </html> | ||
Latest revision as of 13:08, 17 October 2014
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
Figure 1. Bio-Rad agarose gel.
