Register New Projects and Keep Registrations Current/Accurate. The Biosafety Office relies upon new project registrations and Amendments (Administrative/Technical Change), submitted as changes occur, to accurately document work locations, personnel and other aspects of project work. Forms are accessible at: http://www.ehs.ufl.edu/programs/bio/forms/ . UF Policy is as follows.
Register Recombinant DNA (RD) Projects.
For experiments involving the use of recombinant DNA (rDNA)
For experiments involving synthetic nucleic acids that
are designed to integrate into DNA,
are replication competent or able to replicate in a living cell or
code for a vertebrate toxin with an LD50 of <100 nanograms/kilogram body weight.
Register Biological Agent (BA) Projects.
When handling primary human tumor cells or cells immortalized with a virus (e.g., EBV or a retrovirus).
Whenever known or suspected BSL-2 or BSL-3 human, animal, or plant pathogens are handled and do not fall under RD project registration.
Whenever human blood /other tissues known to be positive for any human disease-causing virus or other agent (e.g., HIV) are used in research.
RegisterAcute Toxins (AT) to cover the possession, use, and transfer of biological toxins with a mammalian LD50 of ≤ 100 μg/kg body weight. The AT form must also be completed when organisms, both natural and recombinant, produce these biological toxins.
Register Biological Agents in Storage i.e.any materials described under the BA registration that are not currently being used for experiments and are not part of an active registered project (BA or RD).
Make sure that Appropriate Training has been provided.
Project-specific training must be completed. This is given by the PI or Supervisor; project registration signatures attest to receipt of this training.
Whenever applicable ‘Bloodborne Pathogens (BBP)’, ‘Biomedical Waste’ trainings are required annually and ‘Shipping and Transport of Biological Materials’ every two years. The trainings are available online. See registration details and the training list. Please email the Biosafety Office for more information.
Use PPE Properly.
Gloves are for single-use only; discard with other contaminated waste.
Lab coats must be worn when working at BSL-2, and when handling potentially hazardous or pathogenic material. Use disposable lab coats (they may be re-used) or use a commercial laundry service provider to clean lab coats that are worn as PPE. Do not take them home to wash!
Do not wear PPE in public areas.
Do not wear sandals or open-toed shoes in the lab.
Decontaminate work surfaces (at least daily). Freshly-diluted bleach is effective (UV light may not be effective); ethanol removes corrosive residue but evaporates too quickly to be an effective disinfectant.
Handle Biological Waste Properly. See the Packaging PDF for quick reference. We routinely emphasize the following.
Minimize puncture potential of items going into biohazard bags e.g. put wooden picks/pipette tips into a sturdy bag, empty media bottle or box; bundle serological pipettes or similarly package, re-sleeve or otherwise align them prior to placement in the biohazard bag.
Use a small red bag/horizontal containerinside the BSC to collect biowaste prior to removal for discard. This practice best preserves the laminar flow (protects your work as well as staff working in the area, and also promotes containment of contaminated items).
Use a covered plastic/metal waste container lined with the red Fisher/VWR biohazard bag to dispose of biohazardous material, autoclave if necessary and place the bag inside the fiberboard biohazard box lined with the red liner bag (Stericycle) for final disposal. Do not autoclave the liner bag; only inactivated or noninfectious material should be disposed of in these liner bags.
Keep a tray under biowaste bags. Do not place them directly on carts or on the floor. A tray promotes containment of leaks and facilitates clean up.
All cultures, any recombinant DNA (GMOs), and all potentially pathogenic material (including human cells) must be inactivated (e.g., treated with a final concentration of 10% bleach for at least 30 minutes or autoclaved for 60-90 minutes in a regularly-tested autoclave) prior to final disposal.
Use Sharps Containers Appropriately.
Close sharps container when ¾ full. Label with date closed, PI/Supervisor name, room number, phone number, autoclave if contents are infectious/potentially infectious. Sharps containers must be disposed of within 30 days of closing. Dispose into a red liner bag-lined biohazard box.
If ‘soft’ items i.e. items that cannot cut/puncture the skin like paper, gloves, wrappers etc. are discarded in sharps containers, the container should be dated when first such item is placed in it and should be closed and disposed of within 30days of this date.
Use the “Clean Lab Ware” Disposal Policy According to Guidelines: Follow rinsing, packaging (not in red bags / no biohazard symbols!) and disposal instructions for non-contaminated GLASS and PLASTIC.
Keep Open Flames out of Biosafety Cabinets (BSCs) and ‘Clean Benches’. Bunsen burners are not permitted. Alternatives to flame are strongly recommended. If flame is essential, use a device with safety features such as the Burner Touch-O-Matic [Fisher Cat. # NC-0657135].
Always use HEPA Filters on Vacuum Lines to protect the vacuum system. Filters may be ordered through Fisher (e.g. # 09-744-79; 09-744-75; 09-744-76; 09-744-12; SLFG05010; SLFG85010).
Provide double-containment for storage and transport of infectious/potentially infectious material between labs. Use a durable leak-proof container, such as a Tupperware box, decontaminate the exterior and label with a biohazard sticker.
Have a designated Biological Spill Kit. Keep handy where infectious/potentially infectious and recombinant DNA materials are used. Include spill handling procedures.
Cover/replacefabric chairs in laboratories. CDC/NIH requirements state that nonporous surfaces must cover chairs in order to facilitate spill cleanup. While permitted at designated desk areas, torn or fabric chairs must not be used at lab benches. Try simply “reupholstering” with inexpensive vinyl material or use cling / shrink wrap or plastic bags.
Dispose of Ethidium Bromide Waste Correctly: See Electrophoresis Waste. Use a gel bucket provided by UF Hazardous Materials Management (352-392-8400) to store all ethidium bromide-contaminated material or switch to SYBR green (e.g., Invitrogen’s SYBR Safe product). Destained gels can be placed in the regular trash (note: not the biowaste box).
