General Laboratory Safety

Before we started experimenting in the lab, we managed to know the safety level of the laboratory and classified it as Biosafety level 1. Then we read and learnt to apply and exploit the safety protocols, which were ought to be carried out at this level of Biosafety. These protocols included:

• mechanical pipetting
• decontamination of materials prior to disposal, using an autoclave
• personal protective equipment (lab coats, eye protection, and gloves)
• the prohibition of food, drinks, and any smokable materials, etc.

Furthermore, the whole lab team had previously worked in laboratories of the same biosafety level, hence they were highly-experienced and familiar with the work requirements. To ensure that work in this particular lab goes smoothly, the team was introduced to the safety equipment storage place and safety procedure routes.

Safety Protocols and Precautions

• Personal protective equipment: lab coats, eyewear, and enclosed footwear were used to minimize the possibility of skin contact area with harmful substances. Before leaving the lab, the team made sure to disinfect and sterilize their clothing to prevent the contamination of external environment
• Specific safety equipment: the locations and operating procedures of emergency showers and eyewashes, as well as the fire extinguisher and the panic button
• Waste disposal: the team was introduced to autoclaving protocols and specific chemical waste disposal protocols according to the compounds used
• Chemical safety: the locations and appropriate storage methods for both flammable and potentially hazardous chemicals, as well as the required working environment protocols to handle these substances: working in an active fume hood, etc.

Project Specific Laboratory Safety

We also utilized some compounds that required additional safety protocols:

UV light: We used the UV lights both to sterilize our boxes plus equipment and to cut out gels. Our team members wore protective glasses, which fell under the personal protective equipment criteria. Due to very few personal close-up encounters with UV and taken safety precautions, we completely minimized the possible hazards of encountering UV light in the lab.

Ethidium Bromide: This compound is a potent mutagen and is moderately toxic after an acute exposure. Therefore, the lab team took it upon themselves to familiarize with the appropriate procedures that are required to handle cases of spills or acute exposures. These procedures included working in an active fume hood and wearing personal protective equipment (PPE) to minimize the chance of exposure, as well as extensively studying the material safety data sheet (MSDS) for Ethidium Bromide. We disposed of the product in accord with the disposal protocols and the state safety regulations.

Antibiotics: We used a few main antibiotics for selection procedure. According to the material safety data sheet (MSDS) for ampicillin, chloramphenicol, and kanamycin, these compounds are known to have hazardous effects at high concentrations and both acute and chronic exposure to humans via inhalation, ingestion, or contact with eyes. Therefore, the team wore personal protective equipment (PPE) when handling the compound, studied the safety protocols and were equipped with the necessary information to handle spills, acute exposures, and disposed of the product in accord with the disposal protocols as well as the state safety regulations.

Vaginolysin and human blood: While creating SynDrop cell-free system special attention was paid to the safety regulations as human-specific toxin vaginolysin was implemented into our system. Vaginolysin is the human-specific cytotoxin produced by Gardnerella vaginalis and believed to play a critical role in the pathogenesis of bacterial vaginosis and its associated morbidities. In order to check vaginolysin activity experiments with human blood was essential. To ensure that it compiles with safety guidelines we managed to get safety form by iGEM Safety Committee and agreement to use it for experiments with human blood from Vilnius University.

Improved Laboratory Safety

The main idea of SynDrop project was to create a cell-free system for protein research. This system shows potential to be used for more diverse experiments in the lab. As we managed to develop liposomes which can encapsulate IVTT system with required genetic material, one can perform desired research by reducing work with genetically modified organisms. It is advantageous as experiments can be performed without using additional antibiotics. Also, the risk to spread genetically modified organisms diminishes greatly.