Safety

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Prologue

In research, safety is indispensable, including in iGEM projects. The safety procedures and precautions should be identified and taken into account when designing, preparing and conducting experiments. Safety of our team members is always the number one priority. The lab technicians and researchers at the Department of Biotechnology and Biomedicine of the Technical University of Denmark (DTU Bioengineering) helped us carefully plan the safety aspects of our experiments. After their guidance and the approval of our PI and lab technician, we were allowed to conduct our project in their facilities.

Lab Safety

Prior to commencing our wet lab experiments at the university, we had to complete a mandatory safety and risk assessment of experimental work as mandated by the Danish law. This assesment’s primary objective is to establish the safest way to work and ensure the creation of a safe work environment. In addition to reviewing various work safety documents, passing a comprehensive quiz was a prerequisite for gaining access to the DTU laboratories. This quiz served to identify project-related risks and their mitigation strategies in accordance with Danish regulations governing the handling of GMOs. Furthermore, participation in a mandatory lab safety tour with two laboratory technicians was essential. This tour equipped us with practical knowledge of emergency procedures and facility policies. Additionally, prior to using any potentially hazardous equipment in our laboratories, mandatory training sessions were conducted by our laboratory technicians.

Project Design

General microorganism information Escherichia coli is the chassis for our project, where the two specific strains that we utilize are E. coli BL21(DE3) and E. coli DH5a. These two strains are classified as Biosafety Level 1 and are a part of the white list, indicating that they are safe to handle in a standard microbiology laboratory. All strains were obtained through NEB.

Use of harmful reagents and procedures Perfluorooctanioc acid(PFOA) PFOA is a toxic and carcinogenic compound, which is the environmental pollutant that we are trying to detect. PFOA is a perfluoroalkyl substance and a stable chemical that can repel oil and water to provide surface protection of different products. It is a white powder that can be released into water, and it can readily be absorbed and accumulate in the body.

Due to these properties, PFOA was only handled solubilized to avoid inhalation and ingestion. When working with the solution, gloves were worn and experiments were carried out in the biosafety cabinet. Along with these precautions, we made sure to follow special instructions given by the official DTU website, Kemibrug, for identification of chemicals before we used PFOA for any experiments.

SYBR Safe in gels SYBR safe is a weakly mutagenic substance that is used to visualize DNA in gel electrophoresis. Compared to ethidium bromide, it is a safer choice as it causes fewer mutations. In addition to its mutagenic properties, it is a skin and eye irritant and is suspected of causing organ damage.

To minimize the area where SYBR safe was used, we dedicated an area of the lab to this, along with wearing gloves while working with it. Along with these precautions, we ensured adherence to special instructions given by the official DTU website, Kemibrug, for identification of chemicals before use.

Antibiotics Antibiotics were used for selection procedures. We used ampicillin, chloramphenicol, neomycin, kanamycin, tetracycline and anhydrotetracycline, which are hazardous substances. Ampicillin, chloramphenicol, and kanamycin were already in solution at standard concentrations, provided by our supervisors, whereas tetracycline and anhydrotetracycline were powders. The powders were weighed in a fume bench and solubilized. Along with these precautions, we ensured adherence to special instructions given by the official DTU website, Kemibrug, for identification of chemicals before use.

References