Team:Thessaloniki/Safety

Safety

Safety

During our time in the lab (Biosafety Level 1), our primary concern and focus is following the safety and security procedures and rules of the lab we work in, to ensure the safety of the lab members, society and the environment.

We received Safety and Security training by various members of the labs we worked in. We were taught proper lab practice, sterile techniques and physical biosecurity, along with proper waste disposal and emergency procedures. We were instructed on the function and usage of the various equipment in the lab like Biosafety Cabinets, centrifuges and more. Application of the above in our daily lab practices will prevent any organisms to escape the lab.

We had conversations with professors about various Bioethical issues that might have arisen from implications of different projects that were considered. The present project does not pose any major bioethical questions. Furthermore, early on in the process of deciding on a subject, we were taught about data safety and regulations along with dual use research of concern.

Here, we present an overview of such practices:

Potential Dangers Assessment:

In our project, we primarily used E. coli DH5a cells as chassis for our constructs. Additional strains of said organism that we used as chassis include: INVaF’, C43 (DE3), CC118 λ-pir, Xl-1 Blue. We, also, used Vibrio natriegens as part of the collaboration measurements with iGEM Marburg 2018. All of the above organisms are Risk Group 1 according to the DSMZ. We believe that under the application of proper microbial sterile techniques and waste disposal practices, they pose no significant threat to us, the rest of the lab members, or the environment.
We did not use any part from a risk group 3 or higher organism.
We did not fill in any check-in forms since none of our parts met the requirements to do so. When visualizing gels under UV light in the dark room that were meant to be extracted, we used special equipment that protected us from the radiation.

Potentially Harmful Chemicals:

During our time in the lab we handled some potentially dangerous chemicals. Those include:

• Ethidium Bromide (EtBr) is a potentially carcinogenic molecule that we used to stain and visualise DNA in agarose gels. EtBr was stored in tightly sealed tube to avoid spillage, wrapped in tin foil, at 4 degrees centigrade, and was used with caution and care (gloves, lab coat, goggles).
• Antibiotic powders including Chloramphenicol, Kanamycin, Ampicillin, Doxycycline (used as an inducer) when weighed and dissolved in their respective solvent, were handled with care and caution, along with the usage of goggles, masks, lab coat and gloves.

Waste disposal:

All disposables that came in contact with bacteria were first rinsed with diluted bleach and then disposed off in special containers. The process from there was handled by other lab members.
Ethidium Bromide-containing solutions (TAE/TBE buffers after use, and agarose gels) were disposed of in special 5-gallon pails that are then charcoal filtered.