Team:NCTU Formosa/Safety

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Safety

Inside the Lab

Outside the Lab

Safety

Overview

   Our system provides a nice bio-detector, which serves as a convenient tool for detecting mutagens. Since we plan to utilize our product into reality, we must make sure that these gene-modified E. colis will not be spread into the environment. Accordingly, we cut into different aspects to ensure the safety of our product: the training of the experimenter, the level and the rule of the laboratory, and the safety of our final products using in the environment.

Safety Training Inside the Laboratory

   Our experiments were done in the BSL1 laboratory. All of our team members received several training courses and passed exams offered by Laboratory Management System before entering the laboratory. Each member must wear the lab coat, trousers, gloves, and shoes when carrying out experiments. Before and after the experiments, we have to clean the gloves and the experiment table with EtOH. Everybody strictly followed the experimental procedure and never carry anything out of the lab.

   Besides, to make sure the lysate would not contain any alive E. coli, we boiled the lysate for 2 hours after wrapping the Eppendorf that contained 100μL of our modified E. coli with Aluminium foil and put boiled lysate on an LB plate and cultured it at 37℃ for 16 hours. Figure 1 showed the result of the plate and we could see that the boiled lysate had no alive bacteria exist.

Figure 1: Confirm that no BL21 grow on the LB plate after heat sterilization.
Samples are ydfD + quorum sensing gene.
The right plate underwent heat sterilization and the left plate didn't.

Outside the Laboratory
Confirm the Safety of the Bio-detector

   In the future, our modified E. coli will be taken out of the laboratory for outdoor testing. To make sure no bacteria will be sprayed into the environment, bleach water was designed to be added to the space between two walls to prevent modified bacteria from leaking out when encounter with damages.

Bleach Water Sterilization Test

   We mixed 35μL of bleach water and 35μL of our modified E. coli, Rosetta-gami contained insert part, ydfD-QS gene, put them on LB agar plate, and incubated them under 37℃ for 16 hours. The result shows that no alive E. coli exists on the plate.

Figure 2: Confirm that no BL21 grow on the LB plate after bleach water sterilization.
Samples are ydfD + quorum sensing gene.
Bleach water was added to the right plate while the left plate didn't contain bleach water.

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