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From 2013.igem.org

Contents 1 Student and Lab Safety 1.1 Organisms 1.1.1 Our strains: 1.1.2 Cell lines: 1.2 Meeting with lab and Biosafety training 1.3 Chemical and Biological Reagents 1.4 Flammable Reagent and stuff Safety 1.5 Reseacher safety 1.6 Public Safety 1.7 Environment

Student and Lab Safety

Organisms

Many different strains were utilized in our laboratories for the Oncoli experiments.Our choice of bacteria was Escherichia Coli and Bascillus Subtilis.

Our strains:

• Escherichia coli
• NEB 10
• TOP 10
• BL21(DE3)
• Nissle 1917
• Bascillus Subtilis

Cell lines:

• Head and neck cancer cell lines (74A)
• Colon cancer cell lines (HT-29)
• Breast cancer cell lines (MCF7)
• Normal cell lines (HEK293)

None of the organisms utilized in our lab possess the potential to cause disease in individuals. Though, despite having no association with diseases they are treated as potential pathogens, thus a great care was shown to dispose contaminated and non-contaminated equipments in designated bins. Personal protective equipments such as gloves to handle lab apparatus, laboratory coats and goggles were used to prevent contact with bacteria, yeast samples and avoid sources of ultraviolet radiation. Everyday a member was in charge of cleaning and organising the lab.

Standard protocols were used for all types of genetic cloning and maniulation, including transformation, preparing liquid cultures, digestion, electrophoresis and ligation.

Meeting with lab and Biosafety training

Turgut Ozal University shows great importance to provide safety rules and regulations. Within the laboratory environment, eating and drinking is forbidden in order to prevent contamination. All the members were trained for safety regulations of the laboratory as well as toxicity of the chemicals and solution. The Laboratory-Specific biosafety training checklist was followed to ensure all team members were adequately trained.

Chemical and Biological Reagents

Reagents and equipment including solid and liquid bacterial growth media, micropipette, volumetric pipette tips and centrifuge tubes are autoclaved to sterilize (heated to 121 degrees celsius at 15 PSI) both preceding and following use, ensuring there is no threat of waste contamination outside the lab.

Flammable Reagent and stuff Safety

It was obligatory for each team member to complete fire safety training before beginning the lab work. Potentially flammable reagents (e.g. ethanol and isopropyl alcohol) were stored in a flame protected way. Corrosive and Noxious Reagent Safety Hazardous chemicals and solutions such as EtBr are used in some lab procedures such as gel preparation and electrophoresis.

Reseacher safety

However these chemicals and solutions are used according to the safety rules of the laboratory with care and caution. All the members were trained for safety regulations of the laboratory as well as toxicity of the chemicals and solutions before starting the current project.

Public Safety

When released by accident, our parts and materials actually cause no damage to the general public. Due to the inability of E.coli strains, TOP10 and BL21 to survive out of the lab, they cannot pose any risk to the safety and health of the general public.

Environment

E. coli strains TOP10, NEB10, Nissle 1917, BL21 and bascillus strain have very limited ability to survive outside the laboratory; so this makes them unable to survive or disseminate. Therefore, there is no specific environmental risk associated with the E. coli strains. All bacterial wastes are kept in 10% bleaching solution for one day, and then, are autoclaved to be sterilized. Yet, undesired GMOs may achieve ecologically harmful features.