Team:York/PhosphateAssay

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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.