Our Safe Project Design
Our experimental workflow consists of the following steps: PCR, gel electrophoresis, Zymo cleanup (DNA purification for PCR reaction), Golden Gate assembly, heat shock transformation into E. coli Mach1 cells, picking colonies, conducting a plasmid-encoded reporter assay characterized by Kosuri et. al. (2019), miniprep, sequencing, and analyzing our sequencing results. We will perform this with all of our members trained adequately in wetlab techniques and safety precautions.
The strong promoter sequences we constructed could be used to up-regulate hazardous genes in pathogenic E. coli. Also, we will be working primarily with liquids, which are all non-hazardous, but could nonetheless be splashed in a team member's face or eyes. There will also be a fire hazard when conducting sterile work. Our E. coli are lab strains and do not have the adaptations required to survive outside a controlled environment.
Jim Baugh, Olga Draper, Krystyna Kozakiewicz
Biosafety officers at UC Berkeley
Lab Safety Training by the Berkeley Office of Environment, Health & Safety.
Operating under John Christopher Anderson's BUA.
We have taken all of our team members through lab safety training, providing them a tour of the lab and the safety precautions throughout the building, explaining the hazards that are common to our experimentation, and teaching proper pipetting technique. Each of our experiments has an accompanying video and quiz that must be watched and completed before entering the lab, such that students are aware of the protocol they will be performing and any caveats to be considered. We are working in a BSL1 facility with chemicals and strains that are non-hazardous to our team. We have a rigid scheduling system to ensure that the number of people in the lab is limited, and our experimental progress is logged on a private Github repo.
