From 2011.igem.org

Safety

Our project is "Carnivorous E.coli" and we devoted our whole summer to conduct experiments and make preparations for the jamboree. However, never once did we forget about the importance of safety in the lab.

1. Safety in the Lab

Some of the biological reagents and experiments necessary for the project needed safety measures. They were; Ethidium Bromide (EtBr), phenol, chloroform, 3,5-dinitrosalicylic acid, gas burner and ultra violet LED. To make sure they were correctly used, the safe way of conducting the experiments were taught to the person who was going to deal with these possibly hazardous substances beforehand. The equipments and reagents mentioned above were handled with care.

• EtBr　(Ethidium bromide):　EtBr is regarded as a mutagen, carcinogen or teratogen[1]. Lab members were briefed on the possible effects of EtBr. During the experiments, we wore gloves. After usage, the gloves and gel were separately disposed.

• Phenol and 3,5-Dinitrosalicylic Acid: Phenol and its vapors are corrosive to the eyes, the skin, and the respiratory tract　[2]. 3,5-Dinitrosalicylic acid can cause serious irritation to the eyes[3]. These were handled under the draft chamber.

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

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

• Ultra Violet LED: Ultra violet (UVB) is harmful to eyes. Hence, the LEDs were lit only inside the cardboard box and we only look at it to check if it is properly lighted to avoid long-time exposure.

2. Safety of the General Public and Environment

Not only do we focus on the "safety in the lab", we also had to take care to make sure that the hazardous materials were safely kept inside the lab. We stayed in the lab on a rotating basis to keep out outsiders and thieves, and the door was 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 were only allowed in the BSL1, all bacteria we cultured were categorized in the BSL1 and of minimal safety risks in itself. In the second stage of the brainstorming for our project, we consulted our advisors on environmental risks. The genes we transferred would not be pathogenic, infectious or toxic even if they are misused for malicious purpose. We autoclaved all tips and pipettes which came into contact with the GMOs. Flies we bred would not disrupt 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 experiment plan (the form is at [4] only available to the insiders of the univ.) to the Environment, Safety, and Health Organization, Kyoto Univ. [5] and were allowed to operate genetic modification of the bacteria under the Biosafety level 1. Experiments were planned following the safety guideline of the university [6]. These guidelines are based on national laws and this includes several regulations on GMOs [7]. 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 were separated into 3 groups each supported by one graduate student.

4. Idea

Every team had to check if there were any parts or materials that might raise safety issues. It was, however, really difficult for all the lab members to be aware of it. Our safety motto is "Safety is of primary importance. Discuss with other members everything related to your project". If something like this is written on the Registration page, we believe more people will be aware of the importance of safety. Some might think this is over-protection but like we were thought since we were young "better safe than sorry". This idea can be used for other checkpoints 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.