In this project, our team consistently prioritized biosafety and ethical compliance as the foremost prerequisites. All experiments were conducted within a P1-level laboratory, where the facilities and operational protocols fully adhered to the relevant safety requirements of our institution and national regulations, ensuring the safety of both personnel and the environment. Regarding strain usage, this project employed Escherichia coli DH5α and BL21(DE3):
To ensure safety throughout the entire project lifecycle, we established a comprehensive safety management system. This system encompassed online safety training, pre-experimental safety education, standardized equipment operation procedures, personal protective equipment (PPE) usage, strain management protocols, waste disposal procedures, and contingency plans. Through the strict implementation of operational protocols and waste management measures, we guaranteed the controllability and compliance of all experimental work. This system not only protected the laboratory personnel and the experimental environment but also established a safe and sustainable research foundation for exploring green control strategies for postharvest strawberry diseases.
Under the guidance of our principal investigator, all team members completed an online biosafety course. Through this training, we not only gained a profound understanding of the importance of biosafety and research ethics but also uniformly studied and mastered the Standard Operating Procedures (SOPs) for laboratory equipment, as well as the correct handling methods for chemical and biological waste.
Our experimental work explicitly does not involve any animal or human subjects. All procedures are conducted within the scope of established microbial safety levels and fully comply with iGEM's safety review requirements.
All team members are proficient in accurately identifying various biosafety signs, chemical hazard labels, and equipment status indicators within the laboratory environment. Prior to entering the laboratory, we strictly adhere to regulations by wearing appropriate Personal Protective Equipment (PPE)—including lab coats, safety goggles, and gloves—and conduct mutual checks to ensure proper protection.
Our team has established standardized protocols for the access, storage, and retrieval of biological materials (such as plasmids and strains) before and after experiments, with detailed procedures to prevent leakage and cross-contamination. Additionally, we familiarized ourselves with the laboratory layout in advance, identifying the locations and correct operating procedures for safety equipment including hazardous chemical cabinets, emergency showers, and eyewash stations, ensuring prompt response when needed.
Our team has established data management and confidentiality agreements, committing to ensuring the authenticity and integrity of data throughout the experimental process and strictly protecting any potentially involved personal privacy. We recognize that adhering to safety protocols is not only an act of self-protection but also a reflection of our responsibility to the team and society.
Prior to entering the laboratory, we strictly adhered to safety preparation protocols: under the instructor's supervision, all personnel properly donned lab coats, masks, and gloves to effectively isolate corrosive and hazardous substances. Laboratory personnel were also required to study emergency procedures for chemical exposure and the operation of emergency showers in advance, enhancing their capability to respond to unexpected incidents. During experiments, we strictly prohibited eating, drinking, smoking, or applying cosmetics in the operational areas. Direct contact with broken skin during procedures was forbidden, and strains were neither removed from the laboratory nor subjected to rapid movement. Furthermore, handling unidentified reagents was strictly prohibited, and all instrument operations rigorously followed safety guidelines. For example, centrifuges required symmetrical balancing before initiation and could only be opened after coming to a complete stop. Centrifugation operations mandated the use of sealed rotors or buckets, followed by a 30-second settling period before opening; any tube breakage necessitated immediate handling of fragments with forceps. During pipetting, mechanical pipettors were exclusively used, with containers filled to no more than two-thirds capacity, and direct bodily contact with pipettes was strictly forbidden. For autoclaving, we ensured verification of seal integrity and water levels, limited the load to two-thirds capacity, and maintained continuous supervision throughout the process. Materials were only retrieved using heat-resistant gloves after pressure had normalized and the temperature dropped below 80°C. Biological safety cabinets underwent 30-minute UV decontamination prior to use, maintained standard sash height during operations with restricted working range, received comprehensive disinfection after use, and continued airflow for 10 minutes post-operation. In electrophoresis procedures, we ensured complete gel submersion in buffer, correct electrode connection, strict control of sample loading and voltage parameters, and immediate power disconnection with prompt gel disposal upon completion.
Following experiments, we promptly cleaned and returned all equipment to designated locations, stored reagents and consumables in sealed containers, and meticulously documented equipment usage. Through systematic implementation of comprehensive safety controls spanning personal protection to equipment operation, we effectively mitigated potential safety hazards, ensured secure equipment operation and data reliability, thereby maintaining sustained efficient laboratory functioning.
Within our laboratory safety framework, personal protection is consistently prioritized as a primary objective. All team members have undergone systematic safety training and continuously implement protective measures: strict hand disinfection is performed before experiments; proper attire—including long pants, closed-toe shoes, and tied-back hair—is mandatory; lab coats and gloves are worn throughout all procedures to prevent skin contact with hazardous materials; and consumption of food or beverages in laboratory areas is strictly prohibited. During operations, we adhere to standardized protocols: precise aspiration of supernatants during plasmid extraction to avoid splashing; strict sealing during sterilization procedures to prevent contamination; and careful handling of delicate equipment such as pipettes and centrifuge tubes. Regarding equipment and environmental management, we operate autoclaves in accordance with established guidelines to mitigate high-temperature and high-pressure risks; UV-protective goggles are worn during electrophoresis; and a categorized disposal process is implemented for microbial waste and pipette tips to comprehensively prevent biological contamination. By integrating protective awareness into every operational detail, we have established a multi-dimensional safety defense system encompassing personnel, equipment, and environment.
Our laboratory strictly enforces facility and environmental management requirements, ensuring clear physical separation into clean, semi-contaminated, and contaminated zones to prevent cross-contamination. For routine operations, all work surfaces and equipment are decontaminated using chlorine-based disinfectants or 75% ethanol. Biological safety cabinets undergo thorough disinfection after each use, and any strain spills are immediately contained with chlorine-based disinfectants. All personnel must complete biosafety level training and obtain operational authorization before conducting experiments.
For BSL-1 experiments, researchers are required to wear appropriate personal protective equipment (PPE) including lab coats, disposable gloves, safety goggles or face shields, and non-slip closed-toe shoes. PPE must be removed following a strict "contaminated-to-clean" zone sequence.
To ensure full traceability of microbial strains, we have implemented an electronic tracking system that documents the entire lifecycle from receipt and storage to usage and disposal, with all steps verified by dual personnel. All procedures must comply with established biosafety level requirements. Critical operations require verification of strain identification numbers, species, and concentrations. Lyophilized strains must be reconstituted within biological safety cabinets, and storage follows categorized labeling protocols: short-term storage (4°C) uses sealed partitioned containers, while long-term preservation requires screw-cap tubes with cryoprotection in liquid nitrogen.
All aseptic techniques are performed in biological safety cabinets. A 10-cm radius around alcohol lamp flames is maintained as sterile zones, and inoculation loops must be flame-sterilized and cooled before use.
The waste disposal process requires strict adherence to sterilization and treatment protocols. All infectious waste—including contaminated pipette tips, gloves, and culture dishes—must undergo autoclave sterilization before disposal as general waste. Liquid waste, such as bacterial cultures and washing solutions, must be treated with chlorine-based disinfectants for 30 minutes before discharge through dedicated drainage systems. Sharps, including needles and inoculation loops, must be placed in puncture-resistant containers, which should be sealed and sterilized when 3/4 full, then handed over to professional agencies for recycling and disposal. All liquid microbial waste, including bacterial, yeast, and mold cultures, must undergo high-pressure steam sterilization to ensure complete control of biosafety risks throughout the process.
In the comprehensive management of laboratory waste, we rigorously implement standardized operating procedures: during transportation, leak-proof dedicated carts are used, with operators wearing gloves and protective goggles, following designated routes and schedules to avoid peak hours when moving waste to centralized collection points; upon receipt, container integrity and label completeness are verified, with waste type, quantity, and disposal information recorded. During temporary storage, waste is managed through categorized zoning (chemical waste liquids, biological waste, sharps, etc.), equipped with leak-proof trays, fireproof cabinets, and ventilation systems. The storage area is furnished with absorbent materials, firefighting equipment, and warning signs, ensuring strict separation of incompatible substances and compliance with storage height and capacity regulations. Finally, the EHS department or specialized disposal companies conduct compliant pretreatment, complete hazardous waste transfer manifests, and arrange for qualified units to carry out incineration or landfill disposal. Both parties conduct on-site inventory checks and sign confirmation documents, with all processing records—including registration forms, transfer manifests, and disposal contracts—archived in accordance with legal requirements, achieving fully traceable closed-loop management of waste from generation to final disposal.
Our team has established a tiered emergency response mechanism and developed standardized disposal procedures for potential accidental exposures and leakage incidents during experiments:
All incidents must be registered through the Laboratory Safety Accident Reporting System, with simultaneous root cause analysis to continuously improve emergency protocols.
In experiments involving volatile organic compounds (VOCs), aqueous solutions of the respective compounds were utilized. Acknowledging the potential volatility risks associated with these compounds, all procedures related to the preparation and handling of VOC solutions were conducted within
All antifungal validation experiments employed Saccharomyces cerevisiae as the model fungal organism.
For safety considerations, our hardware demonstrations utilized only the visual reporter system (violacein) and purified enzyme preparations,
Our detection hardware incorporates multiple safety-by-design features:
Throughout this project, our team has consistently prioritized biosafety as the fundamental prerequisite for all experimental work. Through systematic safety training, we have comprehensively mastered standard procedures for personal protection, equipment operation, and emergency response. By establishing an electronic strain traceability system, we have achieved precise end-to-end management from material receipt to final disposal. Through routine measures including daily disinfection, equipment inspection, and categorized waste sterilization, we have implemented safety protocols in every experimental detail. We fully recognize that strict adherence to biosafety regulations is not only essential for personal protection but also represents our commitment to scientific integrity and social responsibility. These efforts have not only ensured the accident-free execution of experiments but have also cultivated our rigorous and truth-seeking scientific attitude.
