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Team:WPI-Worcester/ATF1
From 2014.igem.org
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<p>Our final Experimental procedure combined our best practices across our previous experiments. In the last procedure, we performed 10 different trials 3 times each. The constitutive promoter was used as a theoretical maximum possible yield. from the isoamyl acetate output observed, we used it as the base standard that the other results were based upon. The yields were based upon the Corrected abundance are of the peaks identified through mass spectroscopy as isoamyl acetate. The results of the other trials and their errors were normalized to the value of the constitutive promoter. </p> | <p>Our final Experimental procedure combined our best practices across our previous experiments. In the last procedure, we performed 10 different trials 3 times each. The constitutive promoter was used as a theoretical maximum possible yield. from the isoamyl acetate output observed, we used it as the base standard that the other results were based upon. The yields were based upon the Corrected abundance are of the peaks identified through mass spectroscopy as isoamyl acetate. The results of the other trials and their errors were normalized to the value of the constitutive promoter. </p> | ||
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Revision as of 19:51, 17 October 2014
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Team:WPI-Worcester
From 2014.igem.org
Better Biobrick Characterization
The ATF1 enzyme converts isoamyl alcohol to isoamyl acetate, which gives off a strong banana odor. The conversion of isoamyl alcohol to isoamyl acetate can be monitored using gas chromatography and mass spectroscopy. Thus, the efficiency of the ATF1 enzyme can be characterized.
We have constructed two different biobricks that include the ATF1 gene. The first biobrick, K1423007, is a construct consisting of an arsenic inducible promoter, a ribosome binding site, the ATF1 gene, and a double terminator. The second biobrick, K1423008, consists of a high strength constitutive promoter, a ribosome binding site, the ATF1 gene, and the double terminator.
BBa_K1243007: Inducible Banana Odor Generator
BBa_K1243008: Constitutive Banana Odor Generator
Before we could quantify our results through gas chromatography, we had to make sure that the liquid LB agar that cells were grown would not interfere with our ability to read the presence of isoamyl alcohol and isoamyl acetate. Included below is a chromatograph showing both of these compounds injected into LB form cell growth until at 5mM concentration.
LB solution injected with 5mM isoamyl alcohol and 5mM isoamyl acetate
In order to efficiently quantify the performance of these two constructs, we designed and performed a number of experiments based around E.Coli with the constitutive promoter plasmid, and E.Coli with the arsenic inducible promoter. The full protocol is located in our protocol section, and can be located through this link. Some findings through these experiments revealed that:
Our final Experimental procedure combined our best practices across our previous experiments. In the last procedure, we performed 10 different trials 3 times each. The constitutive promoter was used as a theoretical maximum possible yield. from the isoamyl acetate output observed, we used it as the base standard that the other results were based upon. The yields were based upon the Corrected abundance are of the peaks identified through mass spectroscopy as isoamyl acetate. The results of the other trials and their errors were normalized to the value of the constitutive promoter.
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