Overview
Our team 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.
Lab Safety
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, materials are autoclaved in the autoclave room to prevent contamination and hazard before use.
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.
Project Safety
We used strains of Escherichia coli K12 with various promoters to measure measure the fluorescence response elicited when they interact with several molecules: carbaryl, 3-phenoxybenzoic acid, lovastatin, butanoyl-homoserine lactone, Phenylglyoxylic Acid, propoxur, perfluorooctane sulfonate, cis-naphthalene dihydrodiol, diethyl phthalate, and tartaric acid. The E. coli used was non-pathogenic and posed minimal threat to human safety.
All experiments performed with the E. coli and molecules, including growing the strains, mixing them with the molecules, titrations, and measuring the fluorescence, were performed in a controlled environment under the supervision of experienced scientists. The assays used for these experiments include the LB broth and M9 media with kanamycin our team created using powdered yeast, salt, and water. The only source of contact with the compounds would be accidental contact. The samples were handled in using personal protective equipment like goggles, lab coats, and gloves to further land micropipettes to limit contact. Additionally, any transfer of materials was done using micropipettes for safety. The molecules were all diluted in DMSO, water, or ethanol, further reducing their minimal toxicity and limiting our contact with them.
The molecule-promoter pairs produced a high fluorescence in the initial screening and a measurable signal corresponding to increasing concentrations of the molecule during titration are specifically identified in our results.
References
[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
