Team:Athens/Safety
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Safety
Laboratory safety
Safety was a very important aspect of our wet lab practices from Day one. We underwent thorough Safety Training, which included proper microbiological aseptic technique, waste management, as well as accident management. We were introduced to the proper use of the equipment we were going to be using, and the dangers of various commonly used chemicals were emphasized upon, as well as ways to manage possible accidents and spills.
The laboratory dress code was strictly followed, which included closed shoes, full-length trousers, and mandatory use of a lab coat, not only when doing experiments, but also when spectating. The lab coats were always kept in a drawer located inside the lab, to ensure no contamination. This minimized any risks of hazard to ourselves, and any accidental release of microorganisms in the environment.
We were very particular about our waste management, as this is often a way in which microorganisms or toxic chemicals are released to the environment due to laboratory experimentation. We made sure to never discard any living microorganisms, and all chemicals were discarded according to the manufacturer’s orders, and local regulations.
Microbiology
Microbiological assays played an important role in our very short stay in the lab. This is why we were very particular and proactive regarding such protocols. An aseptic technique was always used, both to ensure the reproducibility of our experiments and the containment of microbiological material. All of our microbiological work was done in a Biosafety Cabinet, equipped with HEPA filters, that ensure no release. All the workstations were sterilized with 70% ethanol both prior to and after each experiment. Nitrile gloves were worn when working with microorganisms.
We were also very particular as to no live microorganism being disposed of improperly. Our used Petri dishes were all autoclaved before discarding, and all liquid cultures were incubated overnight with a strong chlorine/soap solution, before disposal.
Safety also played a vital role in our selection of chassis and organisms used, all of which were deemed BSL Level 1. The main organism used for plasmid propagation was E. coli BL21 (DE3). For the "Improve a part" criterion we used also E. coli DH5-Alpha. These are both very common engineered laboratory microorganisms, with no risk of human pathogenesis.
Molecular Biology
Our project, as any synthetic biology project, requires molecular manipulations that do not come risk-free. Most of the risk harbored by genetically modified organisms is, however, mitigated by our very careful manipulations to ensure No Release. Another risk in molecular microbiology is the use of antibiotics and the genes that confer resistance to them. Special precaution is taken to make sure that all antibiotics used are not the most clinically significant, and also that the transfection with the resistance genes is done in a controlled environment, only to the designated recipient strains.
Project Safety
Our proposed end product will be the lyophilized lysate of the cells or even a purified mixture of the produced dNTPs and Pfu polymerase. This will confirm the biological inactivity of our product, thus making it safe to use. Another possible end product will be the engineered plasmid that can be sent to labs that will develop their own genetically engineered strain. Due to the high dNTP pools, we expect low reproduction of the bacterial culture which means that the genetically engineered strain could not survive outside ideal lab conditions.
Possible end users of our proposed product are researchers and lab personnel that perform PCR or other NAA protocols in multiple settings. These settings include healthcare facilities and diagnostic laboratories, Community Health Centers in outermost regions and island areas, but also academic institutions and High Schools that use reagents for research and educational purposes. As our project is focused on distributing our genetically engineered organisms to these facilities, special precautions should always be taken, since the risk of working with microorganisms is always present. With that in mind, the already mentioned rules must always be closely followed, to ensure safe handling. However, the end-product is not supposed to be used without the controlled supervision of a lab expert, thus the possibility of environmental release is not a concern.