Team:HokkaidoU Japan/Safety

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Safety Questions

Risks to the safety and health of team members or others in the lab

All lab staff is trained according to safety manual provided by Hokkaido University. We took on ourselves to compile a shortlist of often used dangerous materials and safety procedures in our project.

Dangerous chemicals

Chloroform - corrosive and toxic: must be used in fume hood Ethidium Bromide - intercalating agent: must be used with personal safety gear Ethanol - flammable: must not be used near open flame or in large quantities Liquid Nitrogen - cryogenic container and cryogenic gloves must be used

Procedures and equipment

Agarose gel production - heating in sealed container (rupture risk), scalding hot and vicious during preparation (burn injury risk) - remove container lid before heating in microwave, use safety gear, wait for a few moments before removing from microwave Benson burner - fire risk: DO NOT use flammable materials especially ethanol near open fire Centrifuge - high velocity: balance appropriately, observe the machine till it reaches top velocity Autoclave - high pressure: check the water level, DO NOT open when pressurized UV radiation - damage to eyes and skin: use glove and UV box or UV shield

Non-pathogenic bacteria (policy requires treating as pathogenic, as precaution)

JM109 DH5alpha Both of these are lab safe strains. As a precaution all materials coming in contact are sterilized before and after.

Reference Federal Register, (1986) Vol. V1: 88, 6952–16985

Safety equipment

Gloves
Coats
Goggles
UV Box
UV shield

Waste disposal and sterilization

All equipment and waste coming in contact with bacterial is sterilized by autoclave or bleach. All chemicals compounds were disposed according to requirements for their disposal. All table surface used for work were sterilized with 70% ethanol before and after a procedure

Chemical Usage

All chemical compounds were used according to their manuals and respective material safety data sheet

Genetic material

All genes used in this project come from non-pathogenic bacterial strains of E. coli or R. eutropha. Expressed proteins did not show any toxic effect to their host. Our biobricks do not have any foreseeable selective advantage if released to the environment. After consideration we could not find any usage pausing a security concern.

Risks to the safety and health of the general public if released by design or accident.

Also risks to environmental quality if released by design or accident. All material in the lab are kept safe and accessible only to the authorized lab members. All safety precaution were taken for waste disposal. Preventive actions as described previously were taken make sure the bacteria won’t be released into environment. In unlikely event of release bacteria is lab safe and doesn’t have selective advantage to propagate in nature. This bacteria and/or our biobrick are not designed for usage in public.

Risks to security through malicious misuse by individuals, groups or states. And do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?

No biobricks made by us raise any safety issues. As described before all genes used in this project come from non-pathogenic bacterial strains of E. coli or R. eutropha. Expressed proteins did not show any toxic effect to their host. Our biobricks do not have any foreseeable selective advantage if released to the environment. After consideration we could not find any usage pausing a security concern. Bacterial strains are not pathogenic and lab safe.

Is there a local bio safety group, committee, or review board at your institution? If yes, what does your local bio safety group think about your project?

Project conducted at Hokkaido University involving recombination are overseen by officer of "the Safety Office of Genetic Recombination in Hokkaido University". Our lab is equipped appropriately for the manipulation and genetic recombination of bacterial cells. Team members are instructed according to the safety training manual. Permission to conduct our experiments was acquired.

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?

We suggest that users should always check the safety risks and the rules of the parts, devices, and systems. Also local authorities should be consulted. We should always keep in mind to take a look of the safety page related to the parts that you are going to use. To re-check safety issues with professional institutions. Our suggestions are very simple, but as long as you are a scientist and an iGEM members, they could not be forgotten. Talking about benefits of our project, genetically engineered bacteria may accidentally spread out to the environment. If the bacteria could be made aggregate by themselves, it will be easier to collect them. In the future, as a measure to remove bacteria from the environment, study of aggregation protein would be useful.

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