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Team:Macquarie Australia/WetLab/Protocols/SDSPAGE
From 2014.igem.org
(Difference between revisions)
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<div class="cont-out"> | <div class="cont-out"> | ||
| - | <h3>SDS-PAGE</h3> | + | <h3>SDS-PAGE</h3><br/> |
| - | < | + | |
| + | <ul style="list-style-type: decimal;"> | ||
| + | <li>Re-suspend pelleted bacterial cells in 200µL of Milli-Q-water</li> | ||
| + | <li>Transfer 50 µL of suspension into new Eppendorf tubes and combine with 50ul of 2xTruSep sample buffer.</li> | ||
| + | <li>Shear the cells using a Hamilton syringe.</li> | ||
| + | <li>Centrifuge the preparation for 3minutes @ 13,000 rpm.</li> | ||
| + | <li>Load 20 µL of the supernatant into gel.</li> | ||
| + | <li>Conduct electrophoresis at a constant voltage (200V) for 1 hour.</li> | ||
| + | <li>Coommassie Stain for ~30 minutes.</li> | ||
| + | </ul> | ||
| + | |||
</div> | </div> | ||
</section> | </section> | ||
</body> | </body> | ||
</html> | </html> | ||
Revision as of 05:05, 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.
