Bacterial Strains

We chose the bacteria Escherichia coli to conduct our experiments on. The strains that were used were:

• BL21 DE3 Gold
• XL1 Blue
• NEB5 Alfa

All of these strains are classified as Class 1 and therefore follows iGEMs regulations.

Safe Lab Work

When working in a lab, it is crucial to know the risks with the different experiments and bacterias. Therefore, it is important to have a daily routine that every team member follow. Since we only have been working with species from risk group 1, there have not been any major risks.
Our daily routine has involved always wearing lab coats and gloves, and if necessary, we have also used glasses. When working with DNA and toxic chemicals, we also work in a cabinet to make sure that no contamination will occur.

In the case of working with an open flame, no team member were to be left alone with the fire and a set of eyes always paid attention to the potential hazard.

When staining the agarose gel we used Ethidium Bromide (EtBr), which is a dangerous cancerogenic chemical. To minimize the risk we kept the liquid in a container which always has been placed in a cabinet. When in use we open the lid of the container in order to slowly place the agarose gel in EtBr. Immediately afterwards we close the lid and proceed with staining our gel. When working with EtBr it is required to use gloves and after use, the gloves have to be thrown away in a special bin.

During the project, we have also used liquid nitrogen to freeze and lysis our organisms. To prevent any accidents, two of our team members went through an education how to handle this liquid with safety.

Every day after an experiment, we sterilised the lab benches and washed or throw away the used equipment in special waste. No bacteria were ever thrown into the sink because of potential risk of spreading antibiotic resistance. If discarded, bacteria were thrown in a glass container where they remained until the glass container was full. At that point, the container was autoclaved in order to sterile the bacteria-filled liquid.
We have also used three different antibiotics: Ampicilin, Chlorampehicol and Kanamycin. Chloramphenicol and Ampicillin can both be autoclaved and sterile and then flushed down the drain. Since Kanamycin can tolerate heat/autoclaving and have unknown properties it must be submitted for combustion to make it unharmed.

Safe Shipment

When sending our DNA, for sequencing and for DNA Submission for the iGEM-registry we followed all protocols and did not encounter any safety concerns.