1. Lab-Specific Training Personal Protective Equipment –Safety glasses should be worn at all times You might be safe, but how safe is your neighbor??? –Gloves should be worn most times Protection of self from biohazards Protection of experiments from human-hazards –Lab coats provided, but optional Recommend wearing for handling of hazardous materials (e.g. Ethidium Bromide)

2. Good Microbiological Practices Work with live cultures is permissible on open bench (BL1) For sterility –Should be in Biosafety Cabinet, or –Near open flame on benchtop Handle open cultures as little as possible outside of BSC Always wear gloves when working with cultures.

3. Waste Disposal Separate liquid and solid waste Biohazardous Waste –Will comprise most of our waste –Most routine items ( pipets, pipet tips, microfuge tubes, loops, spreaders, agar plates, etc ) will be solid biohazardous waste Dispose of in white cans or small clear benchtop bags Small amounts of liquid okay Clear benchtop bags should be placed in white cans when full.

4. Waste Disposal Biohazardous Waste (cont.) –Gels should be disposed of in the special black container dedicated to gels Other waste should not be placed here. –Liquid biohazardous waste (cultures) must be decontaminated prior to disposal Shake flasks, large numbers of tube cultures, etc. 10% bleach Autoclaving –Decontaminated waste can be sewered (disposed of in the sink).

5. Waste Disposal (cont.) Chemical Waste –Solid chemical waste can be disposed of with biohazardous waste. –Separate basic, acidic, organic, and oxidizing liquid waste (more on this next slide). –Liquid waste containers kept in the fume hood. Mixed Waste –Bio designation takes priority except with volatile organics –May need separate containment for mixed

6. Chemical Waste Four Chemical Waste Containers Alkaline Aqueous Waste – For pH ≥ 7 solutions that are not oxidizing or predominately organic. Acidic Aqueous Waste – For pH ≤ 7 solutions that are not oxidizing or predominately organic. Organic Waste – For waste that is predominantly organic in nature (ethanol, acetone, butanol, etc.), except for waste containing strong oxidizers. Oxidizer Waste – For waste containing any amount of a strong oxidizer (hydrogen peroxide, permanganate, chromates, nitric acid, etc.).

7. Sharps Waste Big red buckets in lab for large sharps –Next to biosafety cabinets –Should mostly be pipets (long plastic ones, tips), syringes Non-sharps should go in biohazard cans Non-hazardous material (e.g. paper towels, printer paper, clean pipet wrappers) should go in standard bins

8. Final Notes To be safe, you must be aware of your surroundings. Assume any new chemical is unsafe until you find out otherwise. Assume any material to be discarded is hazardous unless you know otherwise.

9. Miscellany Storage of chemicals should be in common area, not on personal benches/shelves. Remember to keep orders/packing slips HOUSEKEEPING – more important as lab becomes more crowded.

10. SECURITY The outer lab door should ONLY be unlocked if there are 2+ people in the lab. Both the lab AND office doors should be locked at night. YOUR LAPTOPS MUST BE DOUBLE- LOCKED!!! Laptops should be backed up regularly (register on IS&T website)

11. EHS Rep Weekly Inspection Occurs every Friday by our EHS Rep See level 1 checklist at: http://web.mit.edu/environment/ehs/rep_tools.html http://web.mit.edu/environment/ehs/rep_tools.html

12. Be CAREFUL and use COMMON SENSE! KNOW what you are working with LABEL – especially when using common areas CLEAN up common areas immediately and periodically maintain your lab bench Be RESPECTFUL of other people’s experiments, solutions, reagents, etc.