Difference between revisions of "Team:York/ProjectBreakdown"
Lizalexianu (Talk | contribs) |
Lizalexianu (Talk | contribs) |
||
Line 82: | Line 82: | ||
<div class="section" id="PhosphateAssay"> | <div class="section" id="PhosphateAssay"> | ||
− | + | <h1>Phosphate Assay -Measuring orthophosphate and polyphosphate levels</h1> | |
− | + | <div class="col-md-2"></div> | |
− | + | <div class="col-md-8"> | |
− | + | <p>In order to determine whether our organism "Phil" was accumulating phosphate, we had to develop a test for quantifying the levels of phosphate in media and within our bacterial cells. We developed a reliable technique to measure both orthophosphate and polyphosphate concentrations in cell supernatant and within growth media.</p> | |
+ | |||
+ | <p>Using the <a href="http://www.abcam.com/phosphate-assay-kit-colorimetric-ab65622.html"> ABCAM </a> kit (ab65622) containing malachite green and ammonium molybdate as indicator components, we enhanced the existing protocol to improve the reliability and accuracy of our tests. This was necessary following preliminary experiments on E.coli supernatant showing that the unaltered assay did not detect polyphosphate. Building on multiple tests, we incorporated an acid hydrolysis and neutralisation step in order to accurately measure polyphosphate levels.</p> | ||
+ | </div> | ||
+ | <div class="col-md-2"></div> | ||
+ | </div> | ||
+ | <div class="col-md-1"></div> | ||
</ul> | </ul> | ||
</div> | </div> |
Revision as of 09:49, 16 September 2015
Project Breakdown
Molecular Biology
- Some info
- More info
Phosphate Assay -Measuring orthophosphate and polyphosphate levels
In order to determine whether our organism "Phil" was accumulating phosphate, we had to develop a test for quantifying the levels of phosphate in media and within our bacterial cells. We developed a reliable technique to measure both orthophosphate and polyphosphate concentrations in cell supernatant and within growth media.
Using the ABCAM kit (ab65622) containing malachite green and ammonium molybdate as indicator components, we enhanced the existing protocol to improve the reliability and accuracy of our tests. This was necessary following preliminary experiments on E.coli supernatant showing that the unaltered assay did not detect polyphosphate. Building on multiple tests, we incorporated an acid hydrolysis and neutralisation step in order to accurately measure polyphosphate levels.
Growth Assay
- Some info
- More info