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From 2012.igem.org

Safety questions

Question 1. Would any of your project ideas raise safety issues in terms of:

• researcher safety,
• public safety, or
• environmental safety

In this project two organisms were used, following are the two organisms listed with biosafety level.

Organism	Biosafety
Escherichia Coli	Level 1
Corynebacterium glutamicum	Level 1

Before starting to work in the lab, a risk assessment (developed by the HSE group) was done of all the equipment and procedures to be used in this project. It had to be approved by the HSE group. The grading system used ranges from A to E, where A is no/very low risk, B is low risk, C is moderate risk, D is high risk and E is very high risk. The risk assessment for the project is given below.

Activity	Possible unwanted incident/strain	Probability assessment (1-5)	Consequence, human being (A-E)	Consequence, external enviroment (A-E)	Consequence, eko/material (A-E)	Consequence, reputation (A-E)	Risk value	Comments/status/ Improvement suggestions
Agarose gel electophoresis	Long term health risk from GelRed (unknown effect)	1	Unknown	A	A	B	Unknown	Gelred is said not to penetrate cell membranes, and thus should not act as mutagen even if it is DNA-binding. Gloves also minimize the risk for exposure.
Autoclave	Rapid pressure fall due to opening the autoclave to soon my cause hot liquid burns on eye or skin	2	C	A	A	B	2C	Will not happen if instructions are followed
Bacteria class 1 and recombinant bacteria	Release of GMO to environment. Bacterial infections.	2	A	A	A	B	2A
Bunsen burners	Skin burns	3	B	A	A	A	3B	New rules how to handle burners+ ethanol baths already installed.
Bunsen burners	Initiation of fire	3	C	A	B	A	3C	New rules how to handle burners + ethanol baths already installed.
General lab work	Deviation from established safety rules and routines	--	--	--	--	--	--	No injuries requiring more than simple first aid in these laboratories for the past 5 years, the present rutines seem sufficient
DNA/RNA isolation and purification	Exposure for dangerous for health chemicals: Phenol-chlorophorm mix	2	C	A	A	B	2C	Phenol-chloroform mix requires the work in a ventilation hood only. All waste should be placed in a special box for hazardous materials.
Environmental samples for cultivation and DNA isolation, preparation of	Bacterial infections, production of toxic/infectous compunds from cloned DNA. Usage of hazardous compounds (e.g. chloroform) for DNA isolation.	1	Unknown	A	A	C	1 - Unknown	In general mild injuries, that can be avoided by present rutines and senseable carefulness during lab-work.
Dry ice/etanol	Eye injury. Frostbite	1	B	A	A	B	1B	Eye-protection and gloves for handling the dry ice
Luminometer and fluorometer, use of	Possible exposure health injurious chemicals.	2	A	A	A	A	2A	See MSDS for relevant chemicals
PCR	Exposure to DMSO when used	1	A	A	A	A	1A	PCR-machine should be in ventilation hood when DMSO is added, use lab coat and gloves
pH-adjustments	Exposure to corrosive chemicals: Skin and eye damage	2	B	A	A	B	2B	Eye-protection and lab coat
Sentrifugation (Sorvall + table)	Damage caused by loose rotor	1	C	A	B	C	1C
Supercompetent or electrocompetent cells, making of	Exposure of skin and eyes to extremely cold liquid (nitrogen or ethanol)	2	C	A	A	B	2C	Conducted with extreme caution, and using eye-protection and gloves.
Ventilation hoods, use of	Exposure to hazardous chemicals due to unsufficient airflow (effects on local hood or other hoods)	1	C	A	A	B	1C	Can be prevented by maintaining sufficient airflow
Waterbaths and shaking waterbaths, (heat and water quality)	Electrical hazards, Burns	1	B	B	A	A	1B	Regular service by trained personnel. Heat-proof gloves must be used during handling of hot items. Hot surfaces must be signed.

All of the participants have had previous lab courses. In these courses the participants have learnt about use of safety equipment, first aid, fire extinguishing, proper dispose of waste, and reading and understanding of safety data sheets of the substances in use. Use of different kinds of equipment and methods used have been taught, as well as writing scientific reports. The labs in use in this project are all approved for working with gene modified organisms (GMOs).

Lab coats, safety glasses and disposable gloves are used when necessary and when required by the HSE guidelines and rules. Materials such as restriction enzymes, buffers, PCR and Gel Purification kits are all bought from biotechnology companies. There are none additional safety precautions to be made than the ones already described. When making solutions of antibiotics, gloves and a fume hood are used, in addition to lab coat and glasses.

When the lab’s not in use, it is locked with all of the equipment and materials/organisms safely stored away. A person that doesn’t belong there will not be able to enter. There are also security personel at the school at all times.

Question 2: Do any of the new BioBrick parts (or devices) that you made this year raise safety issues? If yes,

• Did you document these issues in the Registry?
• How did you manage to handle the safety issue?
• How could other teams learn from your experience?

In this project the goal is to get E. coli to react to specific environmental factors that are found around cancer cells, such as high lactate and low oxygen concentrations. These qualities are not connected to the toxicity, infectivity or pathogenicity of the organism and are therefore considered not harmful. The strain of E. coli used is also not pathogenic, the DH5ɑ strain.

Question 3. Is there a local biosafety group, committee, or review board at your institution?

• If yes, what does your local biosafety group think about your project?
• If no, which specific biosafety rules or guidelines do you have to consider in your country?

Question 4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?