Team:Gaston Day School/Project

From 2014.igem.org

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                            The surrounding areas of Duke Energy’s Buck Steam Station have unintentionally been affected with millions of tons of coal ash, containing multiple toxic chemicals including Cadmium. The release of this ash has caused the water to become a high health hazard with the potential to cause a wide range symptoms, from flu-like symptoms to kidney damage, to fragile bones and possibly death, getting progressively worse through prolonged exposure.  As the danger worsens, more lives are affected.
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                          Methods
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In order to minimize the damage caused by cadmium in water, whether locally or globally, our 2012 team created several heavy metal detectors, but in 2013 decided to concentrate on the cadmium detector. The detector responds to the presence of cadmium with green fluorescence. In 2013, the team worked to increase the sensitivity levels of our detector. Our detector needed to be able to respond to cadmium at low enough levels that the detection would be useful, and the presence of cadmium would not already be apparent.This year, we completed the addition of the 2007 Cambridge team's sensitivity tuners to our detector. The sensitivity tuners amplify the signal received by the detector. At the end of last year, after adding the sensitivity tuners, we began to see indications of a peak at lower levels of cadmium than we had previously thought, so to define that peak, we used test points that were closer together. We discovered a peak of fluorescence, and identified our detection points.
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<br>Duplicate Cadmium Experiment
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<br>1. Set up two groups of plastic tubes, with 4 mL of LB in each. The group of tubes that will hold the cadmium detector cells should also contain 4 µl of chloramphenicol.
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<br>2. Add 100 µl of Cadmium detector cells to the tubes that contain chloramphenicol.
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<br>3. Add 100 µl E. Coli to the other tubes.
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<br>4. Add Cadmium to all tubes according to this chart:
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<br>5. Keep in the incubator overnight
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<br>6. Spin the tubes in the centrifuge for five minutes.
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<br>7. Pour off the supernatant.
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<br>8. Resuspend in 1 mL of PBS.
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Revision as of 19:56, 16 October 2014

2014 GDS iGEM Project Page

>Project

The rising cost of petroleum, coupled with the environmental concerns that go with its use, have led to a recent increase in biofuel research. With this research has come developments in using bacteria to produce biofuels such as isobutanol and ethanol. These alcohols are favored because they can easily be swapped into our current infrastructure of car and truck engines. At Gaston Day School, we have decided to launch a biofuel-focused project. To create the alcohols, we are developing alcohol-resistant strains of E. coli through artificial selection. Also, we are using PCR to amplify and ligate the genes GlmZ, GlmY, and IlmV, which are used in native alcohol production. The combination of these genes and the alcohol resistant strains are the first steps in our new biofuels project.

Methods
Duplicate Cadmium Experiment
1. Set up two groups of plastic tubes, with 4 mL of LB in each. The group of tubes that will hold the cadmium detector cells should also contain 4 µl of chloramphenicol.
2. Add 100 µl of Cadmium detector cells to the tubes that contain chloramphenicol.
3. Add 100 µl E. Coli to the other tubes.
4. Add Cadmium to all tubes according to this chart:
5. Keep in the incubator overnight
6. Spin the tubes in the centrifuge for five minutes.
7. Pour off the supernatant.
8. Resuspend in 1 mL of PBS.

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