Safety

Safety is our utmost priority. A collection of strict protocols was installed to minimise risks when conducting the various experiments.

Potential Hazards of Our Experiments

We utilised live bacteria, E. coli, for the cloning of our plasmid. When handling potential biological hazards, personnel are required to wear gloves to ensure safety in the process of conducting the experiments in order to prevent accidents, infections, and contamination. Therefore, necessary safety equipment and safety protocols must be used and followed.

1. Miniprep

Figure 1. Miniprep process

We utilised the Monarch® Spin Plasmid Miniprep Kit for the plasmid extraction of E. coli.

2. Gel Extraction

Figure 2. Gel extraction process

We used Monarch® Spin DNA Gel Extraction Kit for the gel extraction and DNA purification process.

3. DNA Stain

Figure 3. DNA staining in gel electrophoresis

We utilized DNA dyes to visualize the DNA in gel electrophoresis.

Lab Safety Measures

Several safety measures are strictly enforced in order to create a safe space while conducting experiments in the laboratory. As E. coli was handled during our experiment, our laboratory met Biosafety Level 1 with the following safety protocol:

1. Protective Equipment

Figure 4. Personal protective equipment

The use of personal protective equipment (PPE), such as goggles, gloves, and a lab coat or gown. The above was worn when undergoing any experiments and handling chemicals in order to prevent unwanted skin exposure to substances.

2. Daily Decontamination of All Work Surfaces

Figure 5. Decontamination of work surfaces

All workbenches are required to be sanitised with alcohol frequently, to maintain hygiene when conducting experiments, as well as eliminating excess bacteria that could be left over after an experiment, preventing exposure of personnel to potential biological hazards. Immediate decontamination after spills was also in effect.

3. Authorised Supervision

Every experiment conducted in the lab is supervised by qualified teachers and professionals to guarantee a hazard-free environment and secure adherence to safety protocols.

4. Biological Waste Disposal

Figure 6. Biological waste disposal

Every piece of equipment in contact with dangerous chemicals used within the experiments is exhausted into a specialised bin, which is handled with caution and sanitised fully before being disposed of to prevent any chances of a biological endangerment to personnel or any civilians. Infectious materials should be decontaminated before disposal through the use of an autoclave.

5. Personal Hygiene and Safety

Eating and drinking are prohibited in laboratory environments in order to avoid contamination of test samples and work areas. Regular handwashing ensures personal hygiene of personnel.

6. Mechanical Pipetting

All pipetting done in the laboratory for experiments was done with sanitized tips, no mouth pipetting allowed.

Safety of ColoTrace

To provide a safe alternative option for CRC detection, ColoTrace utilises toehold switches due to their non-invasive nature. The assay operates entirely in vitro plus IVE cell-free expression, which eliminates any biohazard risk. The production of the kit is straightforward. Once the toehold switch is ligated to the plasmid, as the plasmid is inserted into the E. coli, then the E. coli would reproduce exponentially with the plasmid along with the toehold switch. After that, a simple translation is done, and the E. coli is then used for cloning in the test kit.

Figure 7. Fume cupboard