Team:TUDelft/SafetyTest
General Safety
The laboratory work of our project is carried out in ML-1 zone workspaces at the Bionanoscience department of the Faculty of Applied Sciences of our university, Delft University of Technology. ML-1 laboratories, comparable to BSL-1 workspaces, are considered as the lowest level of microbiology laboratory biosecurity where we can only make use of non-pathogenic microorganisms. However, we still have strict rules for ML-1 workspaces. Before we were allowed to enter the lab, all team members had to pass the mandatory safety tests to ensure the knowledge of the safety rules and emergency operation procedures. These safety tests consist of the following:
- General building safety (meeting points, emergency numbers, flight plans, etc.)
- General laboratory safety (chemicals, waste disposal, clothing, safety precautions, etc.)
- Biological safety (ML-1 grade safety, biological waste, safety precautions, etc.)
- Laser safety (general safety precautions)
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Lab Safety Rules
- Lab coats are always mandatory in every laboratory and hallway in ML-1 zone.
- Short pants, skirts or open shoes are not allowed when working in a laboratory .
- No eating and drinking is allowed in the laboratory.
- The use of mobile phones is not allowed in the laboratory (except in case of an emergency).
- No personal items are allowed in the laboratory e.g. bags and jackets.
- Hands should be washed on entering and leaving the laboratory.
- Work clean and tidy.
- The bench should be cleaned with 70% Ethanol before the start and after the end of an experiment.
- You are aware where to find personal protective equipment e.g. hand gloves, safety glasses, eye shower and know how to use them.
- You are aware of the use of a fume hood.
- Store chemicals according to PGS 15. (link)?
- Dispose your chemical waste according to the different waste categories.
- Use cryogenic liquids according the procedure.
- Label your samples (name of experimentalist, substance, date of production, expiration date, permit number if applicable).
- Before working with organisms and cell lines you are obliged to make a Risk Evaluation in the Biosafety module (LabServant).
- A researcher is responsible for an up-to-date registration.
- It is not allowed to work with any biological material without permission of the VM and/or Biological Safety Officer of the Bionanoscience department.
- The lab must be clean and tidy.
- Workbench is cleaned with alcohol before and after handling organisms or cells.
- Take off your lab coat, before washing your hands and when leaving the ML-1 lab.
- Long hair is in a tail and e.g. scarves are in the lab coat.
- No jewelry on hands and wrists; these are extreme infection sources.
General
ML-1
Sci-Phi 29 Safety
General safety is important to work in a responsible manner. But besides general safety there are safety aspects which specifically apply to our project.
Biological Safety
Hosts
To show independence of hosts of our platform, multiple constructs have been tested across multiple
bacterial species. All strains used for these purposes are non-infectious, non-pathogenic and are
scaled under biosafety level 1.
Insert Table 1
Vectors
Our part collection provides an array of modular cloning (MoClo) compatible parts, therefore we cloned
our constructs in the MoClo backbones from the MoClo Toolkit. These backbones contain type specific
overhangs. Furthermore our constructs for Bacillus subtilis were cloned into the MK969 backbone.
Finally expression in Vibrio natriegens was achieved by the use of the pET525Cm-YFP plasmid. All
vectors used are derived from backbones which can be used in ML-I laboratories in accordance to the
biotechnology regulations of biosafety (Link here) for The Netherlands.
Insert Table 2
Inserts
Our part collection provides an array of modular cloning (MoClo) compatible parts that can be used for
predictable expression of genes across different bacterial species. Our collection consists of these
inserts or combinations thereof. To ensure the safety of our project we made sure all of these inserts
are safe to work with in ML-I laberatories.
Insert Table 3
Chemical Safety
Not only biological safety but also chemical safety is an important factor to consider. During our project we worked with carcinogenic chemicals like SYBR Safe and Ethidium Bromide. In order to work with these chemicals possible risks were evaluated, together with the biosafety officer of the Bionanoscience department, prior to working with these chemicals. To safely work with these chemicals appropriate measures were taken. Every piece of equipment that comes in touch with these chemicals are located in a contained, marked off, area, where gloves are used at all times. When excising bands from an agarose gel, under illumination of UV light, protective glasses were worn besides the standard safety measures.
Waste disposal
To complete the circle of safe laboratory practices, we made sure all biological and chemical components are disposed in a responsible and safe manner. These practices apply to make sure that all biological material is contained even after experiments are done. Furthermore chemicals which are used should never be a risk for anybody outside of the lab.
Biological waste
One of the important aspects of working an ML-1 lab is the regulation of biological disposal. Genetically modified organisms are not allowed to be released outside the lab and have to be handled with responsibility. All biological waste has been collected in labeled ML-1 waste bottles which were sterilized by autoclaving. Consumables contaminated with biological waste were disposed in ML-1 waste bins which were also sterilized by autoclaving.
Chemical waste
The disposal of hazardous chemical waste was carried out according to the protocol of the Bionanoscience department. For example we separated SYBR Safe (carcinogenic) waste in a special tank, which was collected by a team of specialists in disposal of hazardous chemical laboratory waste. The contaminated consumables were also disposed in hazardous chemical waste bins.