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− | <p style="font-size:medium">From initial iterations of our <a href="https://2018.igem.org/Team:Newcastle/Modelling/Community" class="black">community model</a>, it became apparent that quantitative data on the growth rates of the bacteria were required in order to inform the model. For this, we observed changes in absorbance at 600 nm over 72 hours of the three nitrogen-fixing bacteria and <i>E. coli</i> in liquid culture at 30 °C using a ThermoFisher Scientific Varioskan LUX Microplate Reader.</p> | + | <p style="font-size:medium">From initial iterations of our <a href="https://2018.igem.org/Team:Newcastle/Modelling/Community" class="black">community model</a>, it became apparent that quantitative data on the growth rates of the bacteria were required in order to inform the model. For this, we observed changes in absorbance at 600 nm over 72 hours of the three nitrogen-fixing bacteria and <i>E. coli</i> in liquid culture at 30 °C using a ThermoFisher Scientific Varioskan LUX Microplate Reader (Figure 2).</p> |
<p style="font-size:medium">The data showed that <i>A. brasilense</i> grew at a slow, steady rate before sharply dying off after approximately 60 hours. The slow growth rate is likely to be because its optimal growth temperature is 37 °C rather than 30 °C. <i>H. seropedicae</i> and <i>A. caulinodans</i> showed very similar growth curves when grown at 30 °C: initial growth rate was very fast and then growth became very slow or static after 20 hours. <i>E. coli</i> grew at a medium pace to begin with and steadily slowed down with time. </p> | <p style="font-size:medium">The data showed that <i>A. brasilense</i> grew at a slow, steady rate before sharply dying off after approximately 60 hours. The slow growth rate is likely to be because its optimal growth temperature is 37 °C rather than 30 °C. <i>H. seropedicae</i> and <i>A. caulinodans</i> showed very similar growth curves when grown at 30 °C: initial growth rate was very fast and then growth became very slow or static after 20 hours. <i>E. coli</i> grew at a medium pace to begin with and steadily slowed down with time. </p> | ||
<img src="https://static.igem.org/mediawiki/2018/0/0c/T--Newcastle--ChemotaxisGrowthCurveGraph.png"> | <img src="https://static.igem.org/mediawiki/2018/0/0c/T--Newcastle--ChemotaxisGrowthCurveGraph.png"> | ||
<p> </p> | <p> </p> | ||
− | <font size="2">Figure 2: Growth curves showing changes in absorbance at 600 nm of <i>E. coli, A. caulinodans, H. seropedicae,</i> and <i> A. brasilense</i> in LB at 30 °C for 72 hours. n=4 replicates, error bars indicate standard error of the mean.</font> | + | <font size="2">Figure 2: Growth curves showing changes in absorbance at 600 nm of <i>E. coli</i>, <i>A. caulinodans</i>, <i>H. seropedicae,</i> and <i> A. brasilense</i> in LB at 30 °C for 72 hours. n=4 replicates, error bars indicate standard error of the mean.</font> |
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<img src="https://static.igem.org/mediawiki/2018/9/99/T--Newcastle--ChemotaxisNaringeninKillCurve2.png"> | <img src="https://static.igem.org/mediawiki/2018/9/99/T--Newcastle--ChemotaxisNaringeninKillCurve2.png"> | ||
− | <font size="2">Figure | + | <font size="2">Figure 3: Optical density at 600 nm wavelength of 4 bacterial species (<i>A. brasilense</i>, <i>A. caulinodans</i>, <i>H. seropedicae</i>, and <i>E. coli</i>) after 24 hours of growth when grown in liquid media (LB) containing different concentrations of naringenin. n=4 replicates, error bars indicate standard error of the mean. </font> |
<p> </p> | <p> </p> | ||
− | <p style="font-size:medium">All species successfully grew in the presence of 0-150 μM naringenin (Figure | + | <p style="font-size:medium">All species successfully grew in the presence of 0-150 μM naringenin (Figure 3). However, it was noted that when the concentration of naringenin exceeded 100 μM, the amount of error also increases. This suggests that naringenin begins to have a greater impact on some, but not all, bacteria in the solution. As such, naringenin concentrations of <100 μM were used as part of subsequent chemotaxis assays to avoid negatively impacting bacterial growth. </p> |
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Revision as of 06:01, 17 October 2018