Safety

Introduction

Our project deals with developing a user-friendly kit for rapidly detecting numerous substances. The detection method relies on the chemotaxis motility of E. coli, where a certain substance either repels or attracts the cells, resulting in clusters that are visible to the naked eye.

When working in the lab, safety was always on the top of our concern. Work safety includes everything from proper equipment such as protective gloves, glasses, lab coat etc., to adequate preparations and proper disposal of chemicals and organisms.

As we are both, working with genetically modified organisms and designing a hardware, safety was exercised on the laboratory scale (personal safety) and on the industrial scale (product safety).

Personal safety:

Working at the IGEM Technion lab:

All members of iGEM have completed safety training, provided by our team instructors. It included all laboratory practices such as the hazards and risks associated with different chemical and biological issues. In addition, we received information about the location of fire alarm pull stations, safety showers, extinguishers and fume hoods.

As part of the project our team primarily worked with the following strains of E-coli: UU1250, Top10, B275∆ ZRAS and B275∆ FILM. Those strains do not exhibit any toxicity and belong to Biosafety level 1, which is the lowest risk level. Furthermore, the fact that they prefer temperatures around 37 °C makes it unlikely that they survive for a long time when accidentally released. In addition, we obeyed to the regulations of the Technion [1] to govern biosafety in the lab.

Minimizing Risks:

To minimize risks and conduct a responsible research certain rules were adapted in the lab:

- Wearing personal protection equipment like lab coats, safety goggles and rubber gloves if needed - Consuming any food or drinks in the lab area is prohibited - Smoking in the lab area is prohibited - Autoclaving all waste that came into contact with biological material is mandatory.

Dangerous Chemicals:

Ethydium Bromide- is a known carcinogen, used for DNA staining in agarose gel. Therefore gel preparation was done in designated marked areas, addition of EtBr was done in a hood, and special emphasis was placed on wearing gloves and lab coats when cutting bands from gel using UV.

Liquid nitrogen- is intensely cold and can cause frostbite or cryogenic burns if it is not used, and handled, properly. Therefore, we always handled liquid nitrogen wearing protective clothing including safety goggles, cryo gloves and tongs.

Product safety

Since the channel inside our chip is exposed to the air and not sealed, the modified bacteria can be spilled out of the chip if the user does not hold it properly. That kind of spillage can cause environmental problems. To solve this problem we can try to look for a material to seal the chip.

To make the final product even safer, we can use a kill switch to minimize the risk of accidental release to the environment. For that purpose, we aim to use the kill switch designed by Berkeley 2008 iGEM Team (BbaK112808). The bacterial solution located inside the chip will contain a certain concentration of IPTG. Since IPTG is not metabolized by E.coli, its concentration remains constant. The kill switch turns on in absence of IPTG. in case of an accidental release of the bacteria to the enviornment, the bacteria will be exposed to low level of IPTG and the death rate will increase.The illustrated circuit can be seen in Figure 1.

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