SAFETY

Before beginning work in the wet lab, all members of our team underwent mandatory training to ensure lab safety and proper hazardous waste management. We completed an online course on Lab Safety for Research Personnel and Hazardous Waste Management, which was developed by the UCSC Environmental Health and Safety department (EH&S). This comprehensive course covered essential topics such as personal protective equipment, safe lab practices, lab organization, and the appropriate handling and disposal of chemicals and biohazards.

Upon successful completion of the online course, we received in-person orientations conducted by our principal investigator, Dr. David Bernick. In order to provide us with adequate protection, EH&S supplied our wet lab with standard white lab coats suitable for BSL1 environments, along with lab safety glasses.

In addition to adhering to proper safety protocols, we implemented a system of weekly lab chores to maintain a clean and organized working environment. We consistently emphasized the importance of keeping bench spaces tidy, and we strictly enforced a policy that prohibited individuals from working alone in the lab.

To ensure our proficiency in operating the autoclave located in the UC Santa Cruz Baskin Engineering building, our principal investigator provided us with hands-on training specific to our lab's autoclave.

Selecting a host organism with a proven track record of safety was a top priority for our team as we embarked on expressing our protein. We opted for Escherichia coli (E. coli), a widely used and well-characterized organism. However, to maintain a contamination-free environment, we implemented stringent handling procedures. Waste generated during the experiments, including E. coli cultures and any associated materials, were promptly and properly disposed of in designated biohazard waste bins. Liquid cultures were discarded responsibly using bleach, ensuring complete inactivation of the bacteria. These measures were taken to prevent any unintended contamination and to maintain the integrity of our experiments. By adhering to these specialized handling protocols, we ensured a safe working environment and reliable results throughout our project.

Selecting a host organism with a proven track record of safety was a top priority for our team as we embarked on expressing our protein. We opted for Escherichia coli (E. coli), a widely used and well-characterized organism. However, to maintain a contamination-free environment, we implemented stringent handling procedures. Waste generated during the experiments, including E. coli cultures and any associated materials, were promptly and properly disposed of in designated biohazard waste bins. Liquid cultures were discarded responsibly using bleach, ensuring complete inactivation of the bacteria. These measures were taken to prevent any unintended contamination and to maintain the integrity of our experiments. By adhering to these specialized handling protocols, we ensured a safe working environment and reliable results throughout our project.

To validate the efficacy of Stealth & Chameleon in improving transformation efficiency in engineered cyanobacteria, Microcystis aeruginiosa (M. aeruginosa) UTEX 2385 strain was utilized. This particular strain natively produces microcystin, a hepatotoxin. Working with this strain of M. aeruginosa poses inhalation and skin contact risks to microcystin.

To maximize the safety of our lab members, we followed appropriate BSL-2 guidelines in our lab. When handling M. aeruginosa, we made use of a biosafety cabinet in a contained level 2 alongside appropriate PPE to prevent accidental exposure to microcystin. M. aeruginosa waste generated was disposed of in biohazard bins and liquid cultures were bleached prior to disposal. Our university's Environmental Health & Safety department was made aware of our work to further ensure the safety of everyone involved and the environment.