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Biology Laboratory (Kimball Hall Room 179) Protocol
Hazardous Level 2

View a PDF Version of the Protocol for printing
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1. Bring the materials/supplies you need to perform the work.

a. Supplies/equipment (and a brief description of capability) that will be on-site at NBTC will be listed on the NBTC website; all else must be brought in by the user. All syringe needles must be provided by the user and cannot be kept at NBTC (the user is responsible for proper use and subsequent disposal into a provided "sharps container").

b. All cell media must be brought in by the user.

2. General

a. Safety glasses, a smock, and nitrile gloves will be worn as a minimum at all times. Upon leaving the lab, the user should hang the smock on a rack inside the BioProcessing Room; do not wear it outside the laboratory.

b. Only stock solutions are allowed into the BioProcessing Room. Any chemistries that need to be prepared should either come with the user already prepared or should be prepared inside the Chemistry Room (177), (adjacent to the BioProcessing Room) and brought to the BioProcessing Room.

c. Centrifuge use

i. Any use of the centrifuge will require documentation that the EHS centrifuge training has been taken prior to use. Centrifuge operation will be interlocked to the computer and access allowed only once such documentation has been shown to NBTC administrators.

ii. The rotor blade must be taken out and wiped down after use with 70% ethanol. Wipe out the interior of the centrifuge as well.

d. There will be a roll of disposable paper covering the workbench; after your work is done, this paper should be disposed of and a new covering of paper should be applied. In general, clean up after yourself.

e. Biological Hood

i. Only one person is allowed in the hood at any one time so as to prevent cross-contamination. Users must sign up for the hood (in addition to the Biology Room).

ii. This is not a chemical exhaust or fume hood; there are HEPA filters for particulate control to provide a sterile environment, but this is all. If you need an exhausted hood, use the NBTC Chemistry Room (Kimball 177) which is adjacent to the Biology Room.

iii. Ensure that the hood air blower is already on; if not, press the toggle switch on (the switch light will come on)

iv. Wipe the hood interior using 70% ethanol.

v. Prior to placing anything inside the hood, wipe anything to be brought into it using 70% ethanol (a plastic bottle will be provided). If the material is bagged, wipe the outside of the bag with ethanol, place the unopened bag inside the hood, then open it inside the hood and remove the material from the bag. Upon exiting the hood, place your material inside the ethanol wiped bag, tape it shut, then remove it from the hood. Take out any other personal tools. Finish with wiping the interior of the hood.

vi. Leave the blower on the hood on after use to ensure continuity of a clean environment

f. All supplies (bottles, tubes, etc.) must be labeled with:

i. User

ii. Material

a. chemical name(s) including media and media base, and
b. concentrations

iii. Date

g. Proper disposal of used supplies

i. All sharps (razor blades, needles, etc.) go into the sharps container

ii. All biological hazards go into the red bag labeled as such

iii. For small instruments, an autoclave unit will be available, but resides in the (adjacent) Chemistry Room.

iv. Cell culture disposal - All flasks, petri dishes, and any other culture containers must be soaked in Clorox bleach for 10', then rinsed prior to disposal in the biohazard waste container.

v. Any liquid wastes are to be treated with 10% Clorox for 10', then rinsed down the drain.

h. Incubators

i. All incubators must have the following information provided (a form will be at the incubators):

a. User Name, Group Name
b. Emergency Contact Information (phone#)
c. Date In
d. Anticipated completion date
e. Type of cell line
f. Whether the cells are "Adherence" or "In Suspension"?
g. Temperature
h. Gas Composition
i. Incubator ID and Tray

ii. The cell lines must be compatible when sharing the incubator; no bacteria, yeast cells, or viruses are to be incubated.

Iii No changes to the operational parameters of the incubator are allowed once another user is already using the system; talk to an NBTC administrator for an alternative incubator or reschedule. Temperature will typically be set at 37 deg C, and 5% CO2 however, this and other operational parameters can be defined to suit the experiment.

Iv NBTC administration reserves the right to throw away anything that is or looks to be contaminated.

3. Operational Procedures

a. Hardcopy documents for all equipment will be available on-site

b. "Mini-specs" which give a brief operational description will also be available for easy reference.

This material is based upon work supported in part by the STC Program of the National Science Foundation under Agreement No. ECS-9876771. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation.