For protein expression, the BL21 strain of Escherichia coli (E. coli) was used. Laboratory strains of E. coli are non-pathogenic to humans, classed as biosafety level one.
All lab work was carried out on the university facilities at the lab that fall under biosafety level 1 class. Before any lab work was started, our team had a safety walkthrough with our PIs, during which laboratory rules were discussed, we were instructed in the use of different emergency tools such as fire extinguisher, fire blanket, and emergency showers as well as how one should handle different chemicals, what personal protective equipment one should use, how to handle the waste.
Daily rules were wearing lab coats and safety goggles, wearing gloves were optional or if needed. With COVID-19 restrictions, it was allowed to be no more than 5 people in the laboratory, keeping social distance as much as possible. It was not allowed to be in the laboratory alone, or work after university facilities are closed. When using a UV-table, special protective headgear was used in addition to a laboratory coat and gloves.
In our laboratory Ethidium Bromide (EtBr) was used for DNA staining, the chemical is considered to be cancerogenic, therefore was handled with great caution. It was only used in a specially designated area with its own fume hood, using double gloves. The leftover EtBr is saved in special flasks before being sent to chemical denaturation.
Adjusting pH was done using a strong acid, therefore it was done in the fume hood and using gloves with high resistance to acids and bases. Any chemical spill was cleaned up. Laboratory benches and used equipment were sterilized after each use.
Bacteria carried plasmids with antibiotic resistance, therefore no bacteria were released into the wild, because of the potential risk of spreading antibiotic resistance. If discarded, bacteria were disposed into a glass container where they remained until the glass container was autoclaved. During experiments, we used two different antibiotics: Chloramphenicol and Kanamycin. Kanamycin was autoclaved and sent to combustion to degrade any leftover antibiotic.
