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Team:HokkaidoU Japan/Safety
From 2013.igem.org
Maestro E.coli
Safety
Basic Safety Questions for iGEM 2013
The chassis organism(s) we are using for this project.
- E.coli(K 12) DH5a
- E.coli(K 12) JM109
Highest Risk Group Listed
This is a list of our new coding regions in our projects.
| Part number | Source of DNA | Species | Risc group | Function |
|---|---|---|---|---|
| BBa_K1084009 | Synthesised, Sigma Alderich | E.coli | 1 | Promoter |
| BBa_K1084010 | Synthesised, Sigma Alderich | E.coli | 1 | Promoter |
| BBa_K1084011 | Synthesised, Sigma Alderich | E.coli | 1 | Promoter |
| BBa_K1084012 | Synthesised, Sigma Alderich | E.coli | 1 | Promoter |
| BBa_K1084013 | Synthesised, Sigma Alderich | E.coli | 1 | Promoter |
| BBa_K1084014 | Synthesised, Sigma Alderich | E.coli | 1 | Promoter |
| BBa_K1084015 | Synthesised, Sigma Alderich | E.coli | 1 | Promoter |
| BBa_K1084101 | Synthesised, Sigma Alderich | E.coli | 1 | RBS |
| BBa_K1084102 | Synthesised, Sigma Alderich | E.coli | 1 | RBS |
| BBa_K1084103 | Synthesised, Sigma Alderich | E.coli | 1 | RBS |
| BBa_K1084104 | Synthesised, Sigma Alderich | E.coli | 1 | RBS |
Description of the biological materials we are using in the lab.
Risks to the safety and health of team members or others working in the lab.
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)
- DH5α
- JM109
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
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 by accident.
Some materials pose risks to the general public. For example, Ethanol is a flammable solution so it must not be used by open fire. Not to release those materials, all lab staff is trained according to safety manual provided by Hokkaido University.
Risks to the environment, if released by design or by accident.
The E. coli strains we use in our lab, are lab sage strains. As a precaution all materials coming in contact are sterilized before and after. Reference Federal Register, (1986) Vol. V1: 88, 6952-16985
Risks to security through malicious misuse by individuals, groups, or countries.
There is no foreseeable risk in misuse of our generated genetic material. Our generated genetic material performs basic functions in biology. However, it is impossible to guard against the incorporation of our parts in malicious settings.
Risks which might arise when our project move from a small-scale lab study to become widely used as a commercial/industrial product.
Our project is about optimizing the expression of genes. Our device does not contain a coding site. Therefore, risk will arise when other users assemble our parts with dangerous coding sites. We have to caution the user when assembling with dangerous coding sites.
Design features to address safety risks.
Our device only contains sequences that regulate the expression of genes. (Promoter, RBS, and terminator) Therefore, our device itself does not contain any safety risks and does not have design feature to address safety risks.
Safety training we received.
We all received a lecture class regarding gene recombination that were held in Hokkaido University. It is based on 'Act on the Conservation and Sustainable Use of Biological Diversity through Regulations on the Use of Living Modified Organisms'.
Biosafety provisions
Link to our institution biosafety guidelines.
Our Institutional Biosafety Committee.
We have a permission to engage in the experiments from the safety officer of genetic recombination of Hokkaido University. All members participating in the experiments are registered with this office. All members are trained according to the safety demands of safety officer of genetic recombination.
Our country’s national biosafety regulations and guidelines
Japan is participating in cartagena act. Please refer to a link below.
http://www.bch.biodic.go.jp/english/cartagena/images/e_cartagena.pdf
Biosafety Level rating of our lab.
Our labs Bio safety level is 2.
The Risk Group of our chassis organisms.
The Risk Group of our chassis organisms is 1.
