Safety is also a crucial aspect of our
project, ensuring the health of all participating members and preventing any potential contamination of the
external environment. Although the DNA sequences and proteins used in our project are non-toxic and
non-pathogenic, we still ensure safety at every stage of the project through training before experiments, strict
laboratory rules, and the proper use of a BSL-2 laboratory
Pre-lab training and weekly meetings
All of our team members have undergone
systematic training on biosafety knowledge, including lab skills training, iGEM biosafety guidelines, and
discussion of project safety (Figure 1). In addition, our weekly meetings emphasize laboratory safety and
biosafety related content.
Figure 1. Pre-lab training
Laboratory regulations
Clothing requirements
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In the laboratory, it is mandatory to wear long-sleeved lab coats, masks, and gloves.
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Wearing shorts, skirts, flip-flops, or sandals is not allowed, and long hair must be tied back.
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Lab coats should not be worn outside the laboratory when leaving
Usage of BSL-2 laboratory
In this year's project, we used the nsp5
sequence from SARS-CoV-2 (NC_045512). Although the plasmid used for expressing nsp5 does not have the capability
to infect humans or cause disease, and nsp5 itself is not directly toxic to humans, we still conducted the
protein
expression and purification in a BSL-2 laboratory for safety reasons.
Experiment materials information:
Strains: we only use E. coli for
experiments including DH5 α and BL21.
Plasmids: pGEX-6P-1, pRSF-DUET1 from Suzhou
Silicon BioScience.
DNA: we ordered our gene materials from
Suzhou Silicon BioScience.
BSL-2 laboratory
Our reagent preparation (such as buffer and
bacterial culture medium preparation) was completed in a BSL-1 laboratory, as mentioned in the safety form.
Other
experimental operations related to SARS-CoV-2 nsp5 were conducted in a BSL-2 laboratory. The BSL-2 public
platform
we utilized meets the biosafety level 2 requirements in terms of spatial environment, equipment configuration,
safety management systems, and waste disposal, providing an appropriate experimental platform for our bacterial
culture, protein expression, purification, and characterization.
The platform is clearly divided into
specific areas: buffer zone, bacterial operation zone, and storage area, effectively minimizing the risk of
cross-contamination due to shared space during experimental operations. Below are the experimental conditions we
used (Figure 2-5):
Figure 2. Bacterial operation zone
Figure 3. Germiculture-box
Figure 4. High speed centrifuge
Figure 5. -80 ℃ fridge(for sample
storage)
Other information
Regarding the use of the SARS-CoV-2 nsp5
sequence, we have submitted the corresponding check-in form and received approval.