Difference between revisions of "Team:Aalto-Helsinki/LabResults"
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<p>Before trying to grow new cells in a reactor, the strain had to be tested with vial-scale cultivations. In this way we could gather information about the induction, cell growth rates in TB-media, and some estimates how much time are needed for propane formation after adding IPTG. At the same time we could practise the usage of gas gromatography/mass spectrometry and make proper standards for propane. Basically, terrific broth (TB) containing induced bacteria would be added into 22ml GC-vials. There the cells are incubated certain time and propane accumulates into the head-space of the vial where the injection and analysis would happen. The <a href="https://static.igem.org/mediawiki/2015/4/4f/Aalto-Helsinki_cultivation_for_GC.pdf" target="_blank">protocol</a> contains more detailed information for the preparation of samples. </p> | <p>Before trying to grow new cells in a reactor, the strain had to be tested with vial-scale cultivations. In this way we could gather information about the induction, cell growth rates in TB-media, and some estimates how much time are needed for propane formation after adding IPTG. At the same time we could practise the usage of gas gromatography/mass spectrometry and make proper standards for propane. Basically, terrific broth (TB) containing induced bacteria would be added into 22ml GC-vials. There the cells are incubated certain time and propane accumulates into the head-space of the vial where the injection and analysis would happen. The <a href="https://static.igem.org/mediawiki/2015/4/4f/Aalto-Helsinki_cultivation_for_GC.pdf" target="_blank">protocol</a> contains more detailed information for the preparation of samples. </p> | ||
− | <p>Two samples were incubated 17 h with 50 RPM shaking in 22 ml vials. Propane concentrations were <b>111 | + | <p>Two samples were incubated 17 h with 50 RPM shaking in 22 ml vials. Propane concentrations were <b>111 μg/l</b> and <b>88 μg/l</b> where the litres were as in the volume of cultivation media. Furthermore, strain's growth rate was significant as the OD600 (1 ml cuvettes) increased from 0.1425 to 0.5212 for 1h 10 min time scale. The rate was assumed to be much lower because TB-media contained four different antibiotics which were needed to keep plasmids inside of bacteria. Details about standards, peaks etc. for GS/MS can be seen in <a href="www.eifileaviela.fi" target="_blank">this file</a>.</p> |
<h3>Continuous production</h3> | <h3>Continuous production</h3> | ||
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− | <p>0.5L chemostat experiment with 44 ml/h flow rate was successful. With 1.0 l/h aeration, the propane content of reactor's gas phase was determined to be <b>4.5-22.7 | + | <p>0.5L chemostat experiment with 44 ml/h flow rate was successful. With 1.0 l/h aeration, the propane content of reactor's gas phase was determined to be <b>4.5-22.7 μg/l</b> with GC/MS. The result were calculated when 0.1-0.5 μg of propane was gathered from reactor's gas exhaust into a 22ml gas chromatography vial. However, these values were below GC-standard line so the accuracy may have decreased. The GC-samples were also gathered after the second steady state was over. Nevertheless, the result proves that propane can be formed industrially with continuous production.</p> |
Revision as of 21:14, 12 September 2015