Team:Edinburgh UG/Safety
Safe Project Design
Whole Organisms: The recipient microorganisms are not pathogenic and are ACDP hazard group 1. We are working with disabled laboratory E. coli strains, such as DH5ɑ and TOP10, which cannot colonise the human intestine. Rhodobacter sphaeroides 2.4.1 is a non-pathogenic photosynthetic organism and cannot colonize the human body. Disabled E. coli strains are not expected to survive in the environment in competition with wild type strains. R. sphaeroides 2.4.1 is a wild type strain and so may be able to survive in the environment. However, as a widely distributed photosynthetic organism, it is not pathogenic and produces no toxins. Therefore, we do not expect our organisms to possess any particular environmentally harmful properties.
Parts: The DNA constructs transformed to the recipient strains will not introduce harmful genes, or gene products that may be of risk to Human Health and Safety. The genes will also offer no survival advantage in the wild. The pathways introduced are not involved with pathogenesis, including the genes from T. vaginalis (ACDP 2), which are for hydrogen production and are not implicated with pathogenicity. Therefore, the constructs will not increase the pathogenic ability of the host.
Vectors: Vectors are transmissible, but carry no dangerous cargo apart from antibiotic resistance such as kanamycin. These are routinely used in labs for selection purposes and resistance determinants for these antibiotics occur naturally in the environment. Resulting genetically modified organisms should pose no greater threat to health and safety than the unmodified host organisms.
Disposal: All strains transformed with plasmids, as well as solids and liquids that were in contact with DNA in general, and constructs carrying the genetic information for antibiotic resistance, will be inactivated and disposed of to mitigate the risk of introducing resistance genes to the environment.
Safe Lab Work: We will not carry out lab work without the supervision of a responsible scientist, and when we are in the lab we will always wear PPE. Liquid and solid wastes will be disposed of by autoclaving according to University standard protocols (121°C for 15 minutes, and 134°C for 5 minutes respectively). Any spillages will be decontaminated with 70% v/v ethanol or 1% v/v Virkon. Razor blades will be used to cut DNA bands out of gels for cloning purposes. Students will be instructed in handling and safe disposal of razor blades and supervised during the procedure. Fume hoods will be operated under supervision when necessary - for example, when working with 2-mercaptoethanol during SDS PAGE analysis.
Hydrogen Risk Assessment: Hydrogen is flammable and can be explosive in a mixture with air and oxygen at high concentrations. Our experiments are highly unlikely to produce large quantities of hydrogen and so should pose no great threat. Our hydrogen collection set up employs the use of barium hydroxide. This is a strong base and is therefore corrosive and toxic. We will be performing experiments using this base within a fume hood to contain the liquid and minimise the toxicity in the laboratory.
Fume Hood in the lab to contain toxic substances during experiments
PPE is provided to each student for wear in the lab
Easily seen and accessible first aid kit
Easily seen and accessible fire blanket
Emergency eye wash clearly signposted and easy to use