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.