Before the team began experimenting, everyone received online education about lab safety. Each member watched videos and passed a small quiz given by our team leader. Some of the educational content is about pipette usage and cell culturing. Following the iGEM Safety Policies, we did not release bacteria into the environment and limited our research to E. coli, a relatively safe bacteria for research purposes. We also have some offline education sessions, involving our biosafety committee. We follow our guidelines about biosafety strictly, and specifics of the guidelines are listed below. Following our guidelines, we are glad to say that we have remained safe throughout our wet lab experiments, as there were no accidents concerning our safety.
Biosafety Committee
We have our own biosafety committee, which consists of two research teachers. They oversee proper work area conditions by checking on the disposal of Petri dishes and liquid wastes, sanitation, etc. Moreover, they offer professional safety training for each and every of our team members. Overall, the teachers make sure we work in a safe environment and that we leave it sanitary after our experiments.
Safety Training
We’ve received related training on topics as follows.
lLab access and rules
lResponsible individuals
lDifferences between biosafety levels
lBiosafety equipment (such as biosafety cabinets)
lGood microbial techniques (such as lab practices)
lDisinfection and sterilization
lEmergency procedures
lTransport rules
lPhysical biosecurity
lPersonnel biosecurity
lDual-use and experiments of concern
lData biosecurity
lChemicals, fire and electrical safety
Work Areas
lOpen bench
Microorganism
Escherichia coli DH5α, DB3.1, BL21
Risk Management Tools
Accident reporting procedure including emergency phone number and the instructor, who is in charge.
Personal protective equipment including lab coats, gloves, eye protection, etc.
Safety Regulations
lAll personnel received a tour around the lab and were informed of the rules upon entering the lab, the main rules include:
lNo running or sudden movements in the lab.
lNo food or dink in the lab.
lAll experiment waste must be thrown in a designated bin, which will then be put through a machine for sterilization.
lAll pipettes must be sanitized after use.
lAll equipment used must follow the user manual.
lExperiments can only be performed with gloves and lab coats on.
lAll personnel must wash their hand upon entry into the laboratory and wear suitable clothing.
Reflection
We are very glad to say that there have not been any accidents or safety hazards happening during our entire lab process. Although there have been ups and downs in our expperiment process, we all made sure to remain safe by following the protocols. In future experiments and projects, we will strive to maintain our overall biosafety.
Reflection for PCR section
In the initial PCR attempt, all three gene fragments (EGFP, ccdB, RIP) were successfully amplified, while the amplification of pTracK99 failed. To identify the issue causing this failure, adjustments are made to the annealing temperature for pTracK99. Instead of the initial 55˚C, two groups of pTracK99 were placed to anneal at 52˚C and 50˚C respectively. After PCR, gel electrophoresis showcases that the group annealed at 50˚C yielded more pTracK99 fragments. From this observation, we reached the conclusion that lowering the annealing temperature is the key to successfully amplifying pTracK99. After further trial and error, we set the annealing temperature for pTracK99 to be at 48˚C.