Team:Macquarie Australia/WetLab/Protocols/SDSPAGE

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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>
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<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Heat">Heat Shock</a></li>
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<li><a href="https://2014.igem.org/Team:Macquarie_Australia/WetLab/Protocols/Operons">Induction of Operons</a></li>
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<h3>SDS-PAGE</h3>
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<h3>SDS-PAGE</h3><br/>
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<p></p>
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<ul style="list-style-type: decimal;">
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<li>Re-suspend pelleted bacterial cells in 200µL of Milli-Q-water</li>
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<li>Transfer 50 µL of suspension into new Eppendorf tubes and combine with 50ul of 2xTruSep sample buffer.</li>
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<li>Shear the cells using a Hamilton syringe.</li>
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<li>Centrifuge the preparation for 3minutes @ 13,000 rpm.</li>
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<li>Load 20 µL of the supernatant into gel.</li>
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<li>Conduct electrophoresis at a constant voltage (200V) for 1 hour.</li>
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<li>Coommassie Stain for ~30 minutes.</li>
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</ul><br/>
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<img src="https://static.igem.org/mediawiki/2014/6/63/Sdspagemq.png" />
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<p><b>Figure 1.</b> Protein bands from SDS-PAGE, visualized under UV light.</p>
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Latest revision as of 13:10, 17 October 2014

SDS-PAGE


  • Re-suspend pelleted bacterial cells in 200µL of Milli-Q-water
  • Transfer 50 µL of suspension into new Eppendorf tubes and combine with 50ul of 2xTruSep sample buffer.
  • Shear the cells using a Hamilton syringe.
  • Centrifuge the preparation for 3minutes @ 13,000 rpm.
  • Load 20 µL of the supernatant into gel.
  • Conduct electrophoresis at a constant voltage (200V) for 1 hour.
  • Coommassie Stain for ~30 minutes.

Figure 1. Protein bands from SDS-PAGE, visualized under UV light.