Safety

1. REGULATORY FRAMEWORK AND OVERSIGHT

Our work follows the Waste Disposal Act (Ministry of Environment, Taiwan) and the Regulations Governing Management of Infectious Biological Materials (Ministry of Health and Welfare, Taiwan). Institutional compliance is ensured through the HIA Science Safety Contract. To maintain alignment with international practices, our principal investigators (PIs) participated in the iGEM-hosted Biosafety GEMinar, and we consulted with Professor Chen at NYCU to ensure the project design meets biosafety standards.

2. LABORATORY ENVIRONMENT AND ACCESS CONTROL

All experiments were performed in a Biosafety Level 1 (BSL-1) laboratory at Hsinchu International Academy. All laboratory work was conducted under the direct supervision of a PI.

3. TRAINING AND RISK AWARENESS

All team members completed an 8-hour project-specific safety training, which included:

- Proper use of PPE (lab coats, gloves, safety goggles, masks).
- Emergency procedures for spills, exposures, or other accidents.
- Waste segregation and sterilization procedures.
- Safe handling and culture of E. coli strains and associated laboratory reagents.
- Proper usage of lab equipment, including pipette, disinfectant bench, autoclave, PCR machine, Bunson Burner, etc.

4. BIOLOGICAL SAFETY

The project investigates the use of Beta-lactamase Inhibitor Protein (BLIP) to resensitize antibiotic-resistant bacteria. To ensure biosafety:

- Only non-pathogenic E. coli strains (DH5α, S17-1, K-12) were used, all classified as Risk Group 1 organisms.
- All genetically modified organisms (GMOs) were confined to the laboratory and sterilized prior to disposal.
- Plasmids containing antibiotic resistance markers were used for selection. All waste was sterilized via autoclaving or chemical decontamination to prevent horizontal gene transfer.

5. CHEMICAL SAFETY

The project involved standard molecular biology reagents, including SYBR Safe DNA stain, agarose, antibiotics (ampicillin, chloramphenicol), sodium hypochlorite (bleach), PBS, TAE buffer components, and glycerol. All chemicals were handled in accordance with institutional safety protocols, and fume hoods were used in any and all lab experiments. Reagents were stored and labeled appropriately, and all hazardous materials were disposed of according to institutional guidelines.

6. WASTE DISPOSAL AND DECONTAMINATION

Laboratory waste management followed Taiwan’s Waste Disposal Act and HIA institutional protocols. All containers used to contain bacteria, broth, or organisms with potential biological hazards were submerged and disinfected in a container bleach under a fume hood.

1. Liquid Waste Disposal
Collect all liquid waste in designated waste containers or bottles (labeled “Biological Waste Liquid”).
Add 10% bleach solution (for example: 100 mL bleach + 900 mL culture medium).
Close the lid, gently mix, and let sit for at least 30 minutes for disinfection.
Dilute thoroughly with water before pouring into the sink for disposal.

2. Solid Waste Disposal
Solid wastes are first chemically decontaminated then placed into designated biological waste bins.
Autoclave: Sterilize the entire bag by autoclaving, then dispose of it as regular trash.

3. Additional Notes
Disinfect the workbench with ethanol after experiments to maintain a clean environment.
At the end of the experiment, wash your hands, tidy up the bench, and put away instruments and consumables.
It is strictly prohibited to discard bacterial samples in regular trash bins or pour them directly into the sink.
For special reagents or waste from genetically modified products, consult your instructor before disposal.

Laboratory waste management followed Taiwan’s Waste Disposal Act and HIA institutional protocols. All containers used to contain bacteria, broth, or organisms with potential biological hazards were submerged and disinfected in a container bleach under a fume hood.

Reference:
Ministry of Environment, Taiwan. (2019) Waste Disposal Act. Retrieved from
https://law.moj.gov.tw/ENG/LawClass/LawAll.aspx?pcode=O0020001

Ministry of Health and Welfare, Taiwan. (2020). Regulations Governing Management of Infectious Biological Materials.
Retrieved from https://law.moj.gov.tw/ENG/LawClass/LawAll.aspx?pcode=L0050024