Team:KIT-Kyoto/Test/Safety/aol

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About Our Lab

  • Does your country use a four-part "Safety Level" rating system for laboratories? (The system might be called in English "Risk Levels", "Bio-Safety Levels", "Containment Levels", "Bio-Security Levels", or some similar name.) If yes, which level is used for the most dangerous organisms?
  • Yes. Level 4 is used for the most dangerous organisms. (True for most countries in Asia, the European Union, and North/South America. This is equivalent to the WHO system.)
  • What is the Safety Level of your lab? (Use the WHO numbering system, in which Level 4 is used for the most dangerous organisms.)
  • Level 1 (low risk, ~= WHO BSL 1)
  • What type of work environments do you use to handle biological materials? Please check all that apply.
  • Laminar flow hood / biosafety cabinet with open front
  • What personal protective equipment do you use in your lab? Please check all that apply.
  • Lab coats Gloves Safety glasses / goggles
  • How do you dispose of biological waste? (For example: liquid cell cultures, agar plates, used pipette tips.)
  • We sterilize it by using the autoclave and send it to the disposal center in our university.


Safety

1. Your Training

  • Have your team members received any safety training yet?
  • Yes, we have already received a safety training.
  • Please briefly describe the topics that you learned about (or will learn about) in your safety training.
  • We have received a lecture by KIT faculty members for half a day. It was about gene operation, gene conservation and the way of disposing the recombinants.
  • Please give a link to the laboratory safety training requirements of your institution (college, university, community lab, etc). Or, if you cannot give a link, briefly describe the requirements.

2. Your Local Rules and Regulations

  • Who is responsible for biological safety at your institution? (You might have an Institutional Biosafety Committee, an Office of Environmental Health and Safety, a single Biosafety Officer, or some other arrangement.) Have you discussed your project with them? Describe any concerns they raised, and any changes you made in your project based on your discussion.
  • Prof. Masanobu Ito is responsible for biological safety. We had a discussion and our plan was accepted, so we did not have to change it.
  • What are the biosafety guidelines of your institution? Please give a link to these guidelines, or briefly describe them if you cannot give a link.
  • In your country, what are the regulations that govern biosafety in research laboratories? Please give a link to these regulations, or briefly describe them if you cannot give a link .

3. The Organisms and Parts that You Use

Species name (including strain) 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?
d-limonene synthase gene(Citrus unshu) 1 NIH Guidenines No risk AB110636/CuMTSE1 Synthesis of d-limonene in Citrus unshiu. Dr. Shimada at NARO(National Agriculture and Food Research Organization)gave it to us. Insert it into Escherichia coli.
gamma-terpinene synthase gene(Citrus unshu) 1 NIH Guidenines No risk AB110639/CuMTS3 Synthesis of gamma-terpinene in Citrus unshiu. Dr. Shimada at NARO(National Agriculture and Food Research Organization) gave it to us. Insert it into Escherichia coli.
beta-pinene synthase gene(Citrus unshu) 1 NIH Guidenines No risk AB110641/CuMTS62 Synthesis of beta-pinene in Citrus unshiu. Dr. Shimada at NARO(National Agriculture and Food Research Organization) gave it to us. Insert it into Escherichia coli.

4. Risks of Your Project Now

  • Risks to the safety and health of team members, or other people working in the lab:
  • Pinene and limonene are mucosal irritative substances, so they may affect people in the lab.
  • Risks to the safety and health of the general public (if any biological materials escaped from your lab):
  • When we show the parts to children who take part in our Open House, they might accidentally lick and intake them.
  • Risks to the environment (from waste disposal, or from materials escaping from your lab):
  • Water may be polluted by waste disposal.
  • Risks to security through malicious mis-use by individuals, groups, or countries:
  • If strangers contaminate the sample, we might be unable to keep it under control.
  • What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)
  • a) Our lab is ventilated continuously.
    b) We warn children not to enter our lab in advance.
    c) We make sure that we do not drain the waste materials and that we bring them to the disposal center.
    d) We make sure to lock the door of our lab when not in use.

5. Risks of Your Project in the Future

  • What new risks might arise from your project's growth? (Consider the categories of risk listed in parts a-d of the previous question: lab workers, the general public, the environment, and malicious mis-uses.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available?
  • Our plan is to make pens with E.coli in ink as a result transduction. If children lick the pen, they might mistakenly intake E.coli.
  • Does your project currently include any design features to reduce risks? Or, if you did all the future work to make your project grow into a popular product, would you plan to design any new features to minimize risks? (For example: auxotrophic chassis, physical containment, etc.) Such features are not required for an iGEM project, but many teams choose to explore them.
  • In order to deactivate E.coli, we grind E.coli containing the parts we developed.

6. Further Comments

  • If you are completing a Preliminary Version of your Safety Form, use this space to describe how far you have progressed in your project, and give some comments about any questions that you left blank.
    You can also use this space for any other comments or additional material.
  • We used PCR to amplify three DNA domains which synthase d-limonene synthase, γ-terpinene synthase, β-pinene synthase respectively. After that we inserted each part into a plasmid DNA.