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Team:Macquarie Australia/WetLab/Protocols/SDSPAGE
From 2014.igem.org
(Difference between revisions)
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<li>Conduct electrophoresis at a constant voltage (200V) for 1 hour.</li> | <li>Conduct electrophoresis at a constant voltage (200V) for 1 hour.</li> | ||
<li>Coommassie Stain for ~30 minutes.</li> | <li>Coommassie Stain for ~30 minutes.</li> | ||
| - | </ul> | + | </ul><br/> |
<img src="https://static.igem.org/mediawiki/2014/6/63/Sdspagemq.png" /> | <img src="https://static.igem.org/mediawiki/2014/6/63/Sdspagemq.png" /> | ||
Revision as of 08:36, 16 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.
