Team:KCL UK/Safety
Safety
As we will be working in a lab full of potential hazards, we took wet lab safety extremely seriously. Before going in we filled out the required safety forms, and our supervisor would then show us all the equipment we may need, and how to operate it safely and efficiently. We were also instructed on basic lab procedures, which shall be listed below. Our aim in the lab was to identify promoters that could produce the correct ratio of viral proteins to make capsids of greater than normal size. However, instead of using viral genes, we instead used GFP to demonstrate the different ratios of product being produced. We used K12 derivatives E.coli X-11 Blue, E.coli DH5alpha and E.coli BL21 as chasses in our project. We would operate on open benches (level 1 lab space).
Organism Safety:
As we were using E.coli, risk group 1 organisms, which have been known to infect the colons of humans, we made sure that the strains we had would not cause any harm to our team working in the lab. We preformed comprehensive literature studies to find if the strains were ever reported as having infected humans. We found that the particular strain of E.coli we used is unable to colonize the human intestinal tract. Furthermore, it is unable to produce the necessary amount of toxin to have an effect on humans. Therefore, we concluded that the strain we were using would be safe to use.
Environmental Safety:
It could be possible that the bacteria we cultured could escape into the surrounding environment, so we also investigated if they could cause harm to other organisms apart from humans. We found that E.coli have no effect on animals, plants or other microorganisms, meaning that it is extremely unlikely that any risks could arise from our experiments.
We used antibiotic resistance to select for bacteria. The bacteria would be resistant to kanamycin, using the psb4k5 plasmid, and chloramphenicol, using the psb1c3 plasmid. All bacterial cultures would be disposed of into separate, specialised waste containers marked for autoclaving.
General Lab Safety:
As we were dealing with risk group 1 organisms, we would follow a biosafety 1 protocol accordingly.
These would include:
- Wearing gloves, goggles, and lab coats
- Using mechanical pipettes
- Using 70% ethanol to clean the workspace before and after experiments
- No phones and computers in the workspace
- No food or drinks in the workspace
- Disposing of sharp objects such as pipette tips into special containers
- Any material in contact with biological parts was disposed of into special bags which would be incinerated
- Appropriate storage of flammable and hazardous material such as ethanol
- Autoclaving material that we used
Equipment
Our lab contained the necessary equipment for any emergenices:
- A fire extinguisher
- A fire blanket
- Eye wash
- First aid kit
Project Specific Safety:
For our project, we had to use certain chemicals and methods that could cause hazards. We sought advice with our supervisor and the Safety Data sheet beforehand. Some of the materials and methods we used were:
- Gel red for DNA staining. We used gel red instead of Ethidium bromide due to it having no mutagenic properties, being unable to penetrate cell membranes and having no toxic effect.
- UV light for viewing DNA bands from gel electrophoresis. We used a screen to see the bands on the gel without being exposed to UV light. This was to avoid retinal damage and blindness from the UV. We would also switch off the light unless necessary to prevent damage to the DNA we were using.
- Using a scalpel to cut out bands from a gel. Our supervisor would demonstrate to us how to use the scalpel safely and effectively, including using ethanol after cutting each band to avoid contamination.
- Gel preparation. We would heat agarose mixed with TAE in a microwave. We would heat for a few seconds before stirring the solution. While stirring we would make sure the flask is pointed away from us in case of any heated liquid erupting outwards.
