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Safety

Safe Project Design

When reviewing the protocols, we considered the risks presented by the organisms and chemicals we used, and we maximized the safety features to ensure lab safety. For the organisms applied in our project, we selected E.coli K-12 strains by considering that the E.coli K-12 strains do not fit into the digestive system in humans and the environment. Additionally, the E.coli phage lambda was also considered a BSL-1 level organism that only infected E.coli but not humans and the environment. We will be cloning anti-lipid A proteins into Lambda Phage. Then we will infect E.coli with it and test the immunogenicity of the cell debris with endotoxin assays such as LAL assay and HEK-Blue cells assay.

While considering the safety of the organisms we applied in the experiments, we also substituted safer materials for dangerous materials in lab protocols. We recognized that gel electrophoresis during these experiments involves ethidium bromide as a dye, which is a carcinogen, and exposure to ethidium bromide is dangerous and poses a risk. Visualization of gels may involve UV, which also offers a level of risk. The SYBR® Safe DNA Gel Stain was then placed because several toxic evaluations showed it had less mutagenic and was safer to work with than ethidium bromide. Moreover, instead of using UV light to visualize and gel extraction, we used a Blue light LED transilluminator which is safer than the traditional UV excitation. The Blue light LED transilluminator reduced the risk of burning skin or eyes and it does not damage the DNA sample we were examining.

Safe Lab Work

Safe lab work was also an important component to ensure our samples were not infected and our students and the environment. Gloves were worn all time while performing any experiments and carrying all samples and organisms. After finishing the lab work of the day, we applied 10% bleach and followed by 70% ethanol to clean up any residue on the lab. Moreover, autoclave and boil methods were also taken into account to work safely and prevent any contamination.