Safety – We collected environmental strains and identified a problem for future iGEM teams that identify unknown strains through 16sRNA.

A large part of our project involved looking to alternative sources for synthetic biology work – non-model strains that behave differently than E. coli that could surpass E. coli in various ways and contribute to new biotechnologies. These non-model organisms may behave better in different environments, handle certain procedures better, or be able to craft larger and more complex proteins than otherwise possible. For them to survive in the environment they must have specialized in some way and our aim was to lay the foundation to tapping into that potential. When we first collected our environmental samples and cultured them to create lysates, a serious question arose: what if these environmental strains we collected from the lake could kill us? This was a very real safety concern that we quickly had to solve. Luckily Sarah Steinbrook, a graduate student that monitored our progress, had encountered this problem before and introduced us to 16S ribosomal RNA gene (rDNA) amplicon analysis, the current standard approach to identifying unknown organisms. A 16S RNA paper

George Wells, a professor of Civil and Environmental Engineering at Northwestern University who also provided us with Nitrosomonas europaea samples, was able to further clarify what to do about potentially harmful strains. If the BSL-2 strain was from a mixed culture, as ours was, then it was likely in low abundance. However, If it was an isolate, he advised that we carefully consider whether or not it would be marginally beneficial to continue working with this strain- and if so, to proceed in a BSL-2 laboratory.