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University of Melbourne
iGEM team

1. Safety Level Rating System

Australia uses a four-part ‘Safety Level’ rating system for laboratories in which Level 4 is used for the most dangerous organisms. Our laboratory is Level 2 (moderate risk).

 

2. Work Environments Used to Handle Biological Materials

Open benches and a laminar flow hood/biosafety cabinet with open front are used when handling biological materials in our laboratory.

Some materials are handled in different work environments, for example DNA gels containing EtBr are only used in designated areas and some procedures involving bacteria that need to be sterile are performed near an open flame, while most other procedures are performed at open benches.

 

3. Personal Protective Equipment

In our laboratory lab coats, gloves and safety glasses/goggles are worn at all times and full face shields are worn when using the UV light box. Latex gloves are used for most procedures in the lab, however, nitrile gloves are worn when handling EtBr.

 

4. Disposal of Biological Waste

Biological waste (e.g. pipette tips) are disposed of in a yellow biohazard which is sealed when not in use. When full, this biohazard bin is sent to be autoclaved before it is disposed with appropriately along with the biological waste from the other laboratories in the institute.

Biological waste such as liquid cell cultures has bleach added to it and sits overnight in the fume cabinet before the liquid is poured down a special biological waste drain and the container is autoclaved.

5. Safety Training

 

The Melbourne iGEM team received safety training provided by The Bio21 Institute. The topics covered in this training included: details of emergency actions, the university OH&S policy and issue resolution procedures, the university's environment policy and procedures, procedures for reporting incidents/near misses, hazards and unsafe acts/work processes, introduction to Bio21 EHS staff, location of relevant EHS documentation, security, 'out of normal hours' access, equipment operating procedures, general safety.

The laboratory safety requirements of the institution can be found here: https://intranet.mdhs.unimelb.edu.au/ohse-training-requirements.


6. Local Rules and Regulations

Christian Rantzau (Bio21 Health & Safety Representative) is responsible for biological safety at our institution. We briefly discussed the program with him and no concerns were raised, nor changes made to the project. He ensured that we were appropriately trained in the safety protocols and procedures of the institution.

The biosafety guidelines of The Bio21 Institute can be found at: http://www.bio21.unimelb.edu.au/bio-21-institute-intranet/environmental-health-and-safety/safety.

The regulations that govern biosafety in research laboratories in Australia may be found via the following link: http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/legislation-2.

7. Organisms and Parts Used

 

Species Name	Risk Group	Risk Group Source	Disease risk to humans?	Part number/name	Natural function of part	How did you acquire it?	How will you use it?
E. coli, BL21(DE3)	1	E. coli K12	Very low virulence			Donation from neighboring lab	This is one of our chassis
E. coli, SHuffle T7 commercially available cells	1	E. coli B	Very low virulence			Acquired from local New England Biolabs agent	This is one of our chassis
				Synthetic peptide 1 based on a de novo amino acid sequence	Not natural	Designed DNA from scratch; DNA synthesised by Life Technologies
				Synthetic peptide 2 based on a de novo amino acid sequence	Not natural	Designed DNA from scratch; DNA synthesised by Life Technologies
				Synthetic peptide 3 based on a de novo amino acid sequence	Not natural	Designed DNA from scratch; DNA synthesised by Genscript
				Synthetic peptide 4 based on a de novo amino acid sequence	Not natural	Designed DNA from scratch; DNA synthesised by Genscript
				Synthetic peptide 5 based on a de novo amino acid sequence	Not natural	Designed DNA from scratch; DNA synthesised by Genscript
				TEV protease gene	Tobacco etch virus protease	Donation from neighboring lab

Risks of Our Project Now

Risk to the Safety and Health of Team Members/People Working in the Lab
E. coli has a low, but not non-existent virulence. For this reason, we wear gloves and lab coats and wash our hands when entering and leaving the labs. When analysing DNA agarose gels both the EtBr solution and the UV light box pose a risk to the health of our team members. In these specific cases we wear nitrile gloves when handling the gel and a face shield when operating the light box.

Risk to the Safety and Health of the General Public
E. coli has a low, but not non-existent virulence. For this reason, we wear gloves and lab coats and wash our hands when entering and leaving the labs. This helps to reduce the risk to the safety and health of the general public.

Risk to the Environment
When disposing of things such as liquid cell culture, we sit them overnight in the fume cabinet with some bleach in the,. In the morning we then pour the liquid down the shoot in the fume cabinet and autoclave the containers. We do not pour hazardous chemicals down the sink. All of these action minimise the risks to the environment.

Risks to Security Through Malicious Mis-Use
Our project does not pose much of a risk in terms of malicious mis-use by others, but to try and prevent this we have key card access into both the building and the labs so that they cannot be accessed by the general public.

Measures Taken to Reduce Risks
We employ safe lab practices (i.e. wearing gloves and lab coats), dispose of waste appropriately, use a baceterium (E. coli) with a very low virulence, and have key card access to the labs.

 

8. Risks of Our Project in the Future

   If our project is successful it will allow people to use the molecules we make as a platform technology for making disulphide bonded peptides. These peptides could potentially be used for harmful purposes. We don’t currently have any design features to reduce the risk of someone using our project for harmful purposes. In the future if it were deemed necessary we could potentially try to limit the types of amino residues or groups that could be added to the arms of our peptide by native chemical ligation.