Our project is centered around an organism that is engineered to express the enzyme nitrogenase in an inducible, oscillatory manner. As a result, we must take safety considerations to protect our team members whilst creating and working with our desired oscillatory organisms. This involves taking into account all BSL-1 safety recommendations, and ensuring our work complies with iGEM rules and regulations.
Our laboratory is classified as a BSL-1 (Biosafety Level 1) laboratory, as well as a standard microbiological lab. Within our laboratory space, we had access to open benches and chemical fume hoods to minimize risks while working with hazardous chemicals. All of the organisms and parts that were used during the course of our project are microorganisms that are on the iGEM White List, and our project did not involve any activities or experiments that are prohibited or considered “unsafe” according to official iGEM guidelines.
Before any experimentation was performed, all members, regardless of whether or not they would be a part of wet lab, were required to do a safety training. These trainings helped us understand the rules and regulations present at Stony Brook University in terms of working in a BSL-1 laboratory. Throughout the period of our experimentation, we worked to follow these safety rules. The major rules followed were: (a) Buddy System – which ensured that at least 2 members were present in the laboratory at all times when wet lab work was run, (b) Careful consideration of risk-averse activities, and (c) Proper following of waste disposal, equipment usage, and pursuing training if we felt we were deficient in any part of our training. Finally, we considered the safety and risk of the work that would be required for our project itself. We ensured that all of the microorganisms or parts we worked with were not infectious or hazardous by themselves.
When considering the design of the project, we made sure that the organisms that we used were not higher than BSL-1. We also performed experiments that were equipped for in our laboratory. If we were ever unsure of some policy, we made sure to ask laboratory technicians in our building, who were always around to help us out.
Prior to starting any lab work, all members of our team underwent safety training that was required by Stony Brook University to work in a BSL-1 laboratory. These training courses covered topics such as working with biological and chemical hazards, and managing the disposal of hazardous waste. In accordance with the safety guidelines mentioned in these trainings, all members wore PPE, including gloves and lab coats, at all times. When necessary, goggles, face shields, and masks were also worn if chemicals posed an extraordinary amount of risk to human health. If a chemical was being used for the first time, the chemical’s Safety Data Sheet was consulted prior to use to determine the necessary level of PPE, as well as proper waste protocols for that chemical. All waste was disposed of according to protocols applying to BSL-1 spaces. For example, all liquid waste was disposed in a separate container under a chemical fume hood, and was labeled with the specific chemicals being disposed.
All living organisms and/or cultures were sterilized prior to disposal (ex. E.coli cultures were sanitized with 10% bleach and left to sit for at least 30 minutes prior to disposal). Throughout the course of our project, many chemicals that pose a hazard to human health were used. This includes chemicals that are classified as carcinogens, highly flammable, and corrosive. Some examples of hazardous chemicals include Acrylamide/Bisacrylamide, Ammonium persulfate, TEMED (1,2-Di(dimethylamino)ethane), Ethidium Bromide, Dimethyl Dulfoxide (DMSO), guanidine HCl, Sodium Dodecyl Sulfate (SDS), and 3-(N-Morpholino)propanesulfonic acid (MOPS). To minimize contact with hazardous chemicals, all lab work involving these chemicals was done in the chemical fume hood.