Team:Macquarie Australia/WetLab/Protocols/SDSPAGE
From 2014.igem.org
(Difference between revisions)
(Created page with "{{Team:Macquarie_Australia/TopNavBar}} <html> <head> <link href="https://2014.igem.org/Template:Team:Macquarie_Australia/css/global?action=raw&ctype=text/css" rel="stylesheet"> <...") |
|||
(4 intermediate revisions not shown) | |||
Line 21: | Line 21: | ||
<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>SDS-PAGE</h3> | + | <h3>SDS-PAGE</h3><br/> |
- | <p></p> | + | |
+ | <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><br/> | ||
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2014/6/63/Sdspagemq.png" /> | ||
+ | <p><b>Figure 1.</b> Protein bands from SDS-PAGE, visualized under UV light.</p> | ||
+ | |||
</div> | </div> | ||
</section> | </section> | ||
</body> | </body> | ||
</html> | </html> |
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.