Team DNHS_SanDiego_CA is in full compliance with iGEM's safety and security guidelines. During the project's design, construction, and implementation phases, our utmost priority has consistently been safety. This commitment extends to incorporating special procedures, practices, and materials as necessary.
All experiments were conducted at Scripps Research Institute in Badran Lab. Our working environment is a standard microbiological lab and is classified as biosafety level 1. There are chairs at each bench, multiple biohazard trash cans, and sinks in every row. At the back of the open bench region are -80 degrees Celsius freezers. Beside the open bench lab space is the incubator room. This room contains a variety of incubators and freezers, along with a fume hood. The neighboring spaces are temperature-regulated rooms that are used to run experiments in hot (37 Celcius) or cold (4 Celcius) environments. Finally, the dark room is used for reading gel electrophoresis results.
All experiments were thoroughly researched and performed with the guidance of scientists at the lab. For the conduction of the experiments, we used open bench areas, sterilized equipment, and biohazard bins to prevent contamination.
Our project exclusively involves activities that adhere to all regulations (Scripps-specific biosafety regulations [1] and California biosafety regulations [2]), and all organisms and components utilized are listed as compliant on the iGEM competition's White List.
We performed titration of our strains with Isopropyl ß-D-1-thiogalactopyranoside (IPTG) and terephthalic acid (TPA) via an assay process. The media being used is Davis Rich Media with carbenicillin and spectinomycin.
Our team designed parts pIGEM1 and pIGEM2, which we will upload to the repository. Throughout the experimentation, we will be using DMSO and ethidium bromide in controlled environments. When transferring the TPA into the tubes for serial dilutions, the TPA was kept in a DMSO solution, so contact with the solution through the transfer process is the only way for accidental contact with the compound. Transfer of Ethidium Bromide from one container to another was performed in a controlled environment with micropipettes to limit safety hazards.
We used an MG1655-derived strain of Escherichia coli in building plasmids using various importers like MucK and TpaK, transcription factors, and promoters. MG1655 is non-pathogenic and poses minimal threat to human safety.
We wanted to address how consumers may feel about having and using a device that contains genetically engineered organisms. Some people may not want to be in close contact with E. coli because they may worry that it would be dangerous to their health. We plan to address this concern by educating those who wish to use this product by explaining that the MG1655-derived strain of E. coli used in this device is not harmful in any way to the human body and will not result in any illnesses. This explanation may be placed on the label of our product device in order for consumers to be aware of their safety. In addition, the early use of our device is marketed towards scientists who are aware of safe lab practices, so they should be aware of the components of our device and possible concerns.
[1] Safety Standards - Scripps Research. (n.d.-a). https://www.scripps.edu/lotz/pdfs/SafetyStandards.pdf
[2] Giso. (n.d.). California code of regulations, Title 8, Section 5193. bloodborne pathogens. https://www.dir.ca.gov/title8/5193.html