Difference between revisions of "Team:UiOslo Norway/Experiments/In vivo Assay Methanol Dehydrogense"

Line 19: Line 19:
 
</ul>
 
</ul>
  
 +
<div class="clear"></div>
 +
<!---------------sponsors------------------------->
 +
 +
<p></br></p>
 +
<div style="float:left; width:100%">
 +
<center><h1>iGEM UiOslo 2015 was sponsored by:</h1>
 +
<hr style="height:2px;border:none;color:black; background-color:black;" />
 +
<img src="https://static.igem.org/mediawiki/2015/0/08/UiOslo_Enova1.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/e/eb/UiOslo_GATC-Biotech.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/4/4a/UiOslo_IDT.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/d/d9/UiOslo_Norwegian_Biochemical_Society.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/f/f0/UiOslo_SnapGene.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/a/ac/UiOslo_Statoil.jpg" Height="60px" hspace="15">
 +
</center>
 +
<hr style="height:2px;border:none;color:black;background-color:black;" />
 +
</div>
 +
 +
<!--------end sponsors---------------------------->
 +
 +
</body>
 
</html>
 
</html>

Revision as of 11:06, 9 September 2015

In vivo Methanol Dehydrogenase Assay

Back to Protocols


  • Inoculate a preculture M9 minimal media containing the appropriate antibiotic and incubate at 37 °C overnight shaking at 220 rpm.

  • Inoculate a mainculture (50 ml) with an OD600 of at least 0.3, add IPTG to a final concentration of 0.1 mM and grow the culture until the stationary phase is reached.

  • Measure OD600 of the culture and harvest a cell pellet of an OD600 of 1 in a volumne of 50 ml by centrifugation at 8000 x g for 10 minutes at roomtemperature

  • Resuspend the cell pellet in 50 ml M9 minimal media and measure the actual OD600

  • Start the experiment by adding 2 ml of 10 % (v/v) methanol

  • Directly after adding the methanol take a 600 µl sample

  • Take a 600 µl sample every 5 minutes over a 1 hour

  • Pellet the harvested cells by centrifugation at 11000 x g for 5 minutes at 4 °C

  • Use 500 µl of the supernatant with 500 µl of Nash reagent and measure absorption at 412 nm and determine the concentration of formaldehyde


iGEM UiOslo 2015 was sponsored by: