We chose to conduct our experiments on the bacteria Escherichia coli. 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 the iGEM regulations.
Safe Lab Work
When working in a lab, it is crucial to know the risks with the different experiments and bacteria. Therefore, it is important to have a daily routine that every team member follows. Since we have only 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 safety glasses. When working with DNA and toxic chemicals, we worked in a fume hood to make sure that no contamination would occur.
In the case of working with an open flame, no team member was to be left alone with the fire.
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 had a permanent place in a fume hood. When in use we open the lid of the container in order to slowly place the agarose gel in the EtBr. Immediately afterwards we closed the lid and proceeded 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 used liquid nitrogen to freeze and lyse our organisms. To prevent any accidents, two of our team members went through an education how to handle this liquid safely.
Every day after an experiment, we sterilised the lab benches and washed the equipment. Waste was thrown in a bin with 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 sterilise the bacteria-filled liquid.
We have also used three different antibiotics: Ampicilin, Chlorampehicol and Kanamycin.
Chloramphenicol and Ampicillin can both be autoclaved 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 harmless.
When sending our DN, for sequencing and for DNA Submission for the iGEM-registry we followed all protocols and did not encounter any safety concerns.
We have also used three different antibiotics: Ampicilin, Chlorampehicol and Kanamycin. Chloramphenicol and Ampicillin can both be autoclaved 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 harmless.