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Summary: Measuring the responsiveness of the serotonin receptor using dynamic range experiment with varying serotonin concentrations.
Documentation: We tested whether the transformed S. cerevisiae cells successfully expressed the serotonin receptor by measuring fluorescence after they were grown in SC media containing 2% galactose and varying concentrations of serotonin. We used a strain of Saccharomyces cerevisiae that had a pAga1 promoter tied to an EGFP reporter. We used an initial S. cerevisiae cell density of OD600 of 0.1 and them in 2% galactose and SC media to induce construct expression. We then allowed our cells to incubate for 2 days at 30°C at 750 rpm, where they grew to an OD600 of around 0.2. We then added varying concentrations of serotonin, ranging from 0.1 µM to 1000 µM (specifically, we tested at 0.1, 1.0, 10.0, 100.0, and 1000.0 µM). We also included a negative control that did not receive any serotonin, and a positive control that had 1 uM alpha factor instead of serotonin (see part BBa_K1829005).
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