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Safety

Our project is "Carnivorous E.coli" and we devote the whole summer for the experiments and preparation for the jamboree. At the lab, however we never forgot the importance of the safety.

1. Safety at the Lab

Some of the biological reagents or experimental necessary for the project need safety concerns. They are; Ethidium Bromide (EtBr), phenol, chloroform, 3,5-dinitrosalicylic acid, gas burner and ultra violet LED. To make sure these are correctly used, safe way of experiments must be taught to the person who deal with these possibly hazardous issues before the experiments.The equipments and reagents above are carefully handled with care.

• EtBr　(Ethidium bromide):　EtBr is regarded as mutagen, carcinogen or teratogen[http://en.wikipedia.org/wiki/Ethidium_bromide]. Lab members are explained

the possible effects of EtBr. So we wear gloves. The gloves and gel are separately disposed.

• Phenol and 3,5-Dinitrosalicylic Acid: Phenol and its vapors are corrosive to the eyes, the skin, and the respiratory tract　[http://en.wikipedia.org/wiki/Phenol]. 3,5-Dinitrosalicylic acid is seriously stimulative to the eyes[http://www.tokyokasei.co.jp/catalog/D0850.html]. These are handled under the draft chamber.

• Chloroform: Chloroform is possibly a carcinogen. To avoid unnecessary exposure, this is also handled under the draft chamber.

• Gas Burner: We use gas burners and heat the air around the workspace to rise, thereby reducing contamination. Although gas burner is a common combustion apparatus we are all familiar with, it can directly cause a massive disaster if used improperly. We make all-out efforts to keep all the flammable items away from the flame and to not pass behind a person who is using a gas burner.

• Ultra Violet LED: Ultra violet (UVB) is harmful to the eye. However, the LEDs are lighted only inside the cardboard box and we have to watch it only when we check if it is properly lighted and long-time exposure to is avoided.

2. Safety of the General Public or Environment

Not only do we note at "safety at the lab", we also have to take care to make sure that the hazardous materials are safely kept inside the lab. We stay at the lab by rotation to keep out outsiders and thieves, or the key is locked and held by the person in charge.

Even when good security is maintained, we should always be careful of misrelease of the bacteria or others(this time: fruit fly). As our experiments are allowed only in the BSL1, all bacteria we culture are categorized in the BSL1 and of minimal safety risks in itself. In the second step of the brainstorming for our project, we consulted the advisors about the environmental risks. The genes we transfer won't raise pathogenicity, infectivity, toxicity when they are misused even on malicious purpose. We autoclave all tips or pipettes that touch the GMOs. Flies we bread don’t disturb the environment because they are the common strain in Japan. Considering these things we concluded there is little risk of environmental disturbance.

3. Regulations

We submitted the experimental plan (the form is at [http://www.esho.kyoto-u.ac.jp/index.php?p=266#format] only available for the insiders of the univ.) to Environment, Safety, and Health Organization, Kyoto Univ. [http://www.kyoto-u.ac.jp/ja/access/faculty/esh.htm] and are allowed to operate genetical modification of bacteria under the Biosafety level 1. Experiments are planned following the safety guideline of the university [www.esho.kyoto-u.ac.jp/wp-content/uploads/2008/05/25_01.pdf]. This guidelines are based on national laws and this includes several regulations on GMOs [http://www.bch.biodic.go.jp/hourei1.html]. We carried out our experiments at the students-laboratory at the Graduate School of Science / Faculty of Science, Kyoto Univ, which has its own department for safety and environment. All of the lab members received training for PCR, culture of cells, miniprep and etc., the minimal genetical operations and usage of autoclave. They are separated into 3 groups each supported by one graduate student.

4. Idea

Every team must check if there is any parts or materials that raise safety issues. It is, however, really difficult for all the lab members to be aware of it. If there is one sentense "Safety is of primary importance. Discuss with other members every thing related to your project" on the Registration page, more people will easily have the chance of becoming aware. Is this over-protection. This idea can be used to other check points such as human practice.

Q1-1. Would the materials used in your project and/or your final product pose: a. Risks to the safety and health of team members or others in the lab? - Yes b. Risks to the safety and health of the general public if released by design or accident? - Yes c. Risks to environmental quality if released by design or accident? - Yes d. Risks to security through malicious misuse by individuals, groups or states? - Yes

Q1-2. Please explain your responses (whether yes or no) to these questions. Specifically, are any parts or devices in your project associated with (or known to cause): - pathogenicity, infectivity, or toxicity? - threats to environmental quality? - security concerns?

A1-2. No

Q2. If your response to any of the questions above is yes: a. Explain how you addressed these issues in project design and while conducting laboratory work. b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry. 3. Under what biosafety provisions will / do you operate? a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible. b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review. c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training. d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.

A2. is at 2. and 3.

Q3. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

A3 is at 4.