Biosafety Level of our Lab

Level 1 – standard microbiological lab 
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Organisms We Used:

Chemicals

Here we present the list of chemicals we used during ourexperiments. We noted down their corresponding hazard levels to ensure that wehandled each chemical safely.

Lab Security Measures

• Handle microorganisms underbiosafety hoods.
• Autoclave the glassware, vessels, tips and media before using.
• Separate trash bins for discarding used plates and culture tubes.
• Separate trash bins for discarding used tips and Eppendorf tubes.
• Clean the biosafety hoods andbenches with ethanol before using.
• Bunsen burners used on benches tohelp avoid contamination.
• Chemicals and reagents are kept in cabinets.  
• Use Virkon aquatic tablets todisinfect the waste tanks.
• Keep the sterile and non-sterile materials separately.

Engineering Safety

To make sure everyone knows how to work safely in the lab, we did the following:

• Lab induction, and safe practices risk assessment
• Emergency Protocols
• COSHH forms: for detergent and use of PET in the imagining suite
• Risk assessments
• Allocation of appropriate personal protective equipment, like lab coats, gloves, and goggles

Biosafety of our parts

We have also taken the safety of our final product into accounts:

• To guarantee that our spores to not contain any foreign DNA, we will use Golden Gate Assembly to assemble self-digesting plasmids, these consist of a reporter protein (GFP), D15exonuclease, Cas9 and gRNA targeting the origin of replication of the plasmid.The D15 sequence comes from E. coli, but will be synthesised and not amplified from genomic DNA. Cas9 assists the cutting of foreign DNA.
• We will also be performing directed evolution of the GerA receptor, which is native to B. subtilis.