As part of our safety training everyone who worked in the lab had to take the following online training tutorials DEHS Introduction: Research Safety, DEHS Chemical Safety, DEHS Chemical Waste Management, and DEHS Globally Harmonized System of Classification and Labeling of Chemicals
To minimize our risk as well as the risk to the environment we chose to work with two non-pathogenic species: Saccharomyces cerevisiae CEN.PK2 and Escherichia coli JM109. In addition to being non-pathogenic, the yeast strain has the additional safeguard in that it is an auxotrophic species unable to synthesize tryptophan, uracil, leucine, and histidine. We always wore nitrile gloves when working on the benchtop, and we autoclaved anything biosafety level 1 organisms could have touched before discarding it as directed by the lab biosafety officer, Chris Flynn. We wore gloves whenever we were working with ethidium bromide, We wore lab coats, gloves, and a face mask whenever we had the UV light on to excise DNA from an electrophoresis gel, and we wore gloves while working in a fume hood when we used hexane.
For our sequences we chose to use sequences all from Biosafety level 1 organisms. In particular we chose to use the Thosea asigna (BS1) viral 2A sequences over the Foot-and-mouth-Disease (BS4) viral 2A sequences just to be on the safe side.