Safety Protocol by HC111213032419

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           Safety Protocol for Safe Handling and Disposal of E. coli
E. coli strains such as K12, DH5, etc. used for the development, propagation and preparation of
recombinant DNA materials. Safe handling of E. coli strains thus needs to be done in accordance
with MUSC policy, the recombinant DNA guidelines and standard microbiological methods.
Laborstory strains of E. coli are considered Risk Group 1 (RG1) agents, which are not associated
with disease in healthy adult humans and require Biosafety Level 1 containment.

1. E. coli strains developed and propagated in this lab will likely contain bacterial vectors with
human or rodent insert genes. Vectors used may include plasmids, bacteriophage lambda,
cosmids, bacterial artificial chromosomes (BACs), P1 artificial chromosomes (PACs), etc.

2. E. coli strains should be handled with appropriate precautions consisting primarily of good
microbiological laboratory techniques. The following precautions should be employed:

A. Access to the laboratory is limited or restricted at the discretion of the laboratory director.
B. Lab personnel must wash their hands after they handle viable materials and animals, after
     removing gloves, and before leaving the laboratory or animal facility.
C. NO eating, drinking, smoking, handling contact lenses, applying cosmetics, etc. in the lab.
D. Do not store food in lab.
E. Never mouth pipette.
F. Sharps should be handled with extreme caution to avoid cuts or autoinoculation during use and
disposal.
         Needles should not be bent, sheared, or recapped. The needle and syringe should be
         promptly placed in a puncture-resistant container and decontaminated, by autoclaving or
         incineration.
G. Minimize splashes and aerosols.
H. Dispose of solid wastes in in orange bags, which are autoclaved and placed in a red biohazard
bag for final disposal.
I. Materials to be decontaminated outside the lab must be placed in a durable leak proof container
and secured for transport
J. Infectious or biohazardous materials must be transported in sealed primary container inside a
sealed durable and leak proof secondary containment labeled with a biohazard sticker.
K. Decontaminate surfaces with 70% ethanol or 10% bleach (made fresh every two weeks)
after a spill or when work is completed for the day.
L. Decontaminate cultures and liquid waste using a final concentration of 10% bleach or 70%
ethanol for a minimum of 20 minutes.
If working with a flame, be sure to keep any ethanol solutions away from the flame at all times.
M. Ensure that laboratory personnel are trained, with signed copies of the safety protocol in the
lab’s safety manual.

3. Safety equipment.
A. Wear lab coats and gloves when working with bacterial cultures.
B. Wear safety glasses when splashes sprays or aerosols can be expected.
C. Dispose of contaminated gloves in Red biohazard bags/containers.
E. No personal protective equipment (PPE) is to be worn outside of the lab.

4. Emergency procedures
        A. Spills of Bacterial Cultures: Notify workers in the area. If aerosol was generated,
leave area for 15 minutes to allow aerosol to settle. Mix spill with bleach to get 10% final
concentration, allow decontamination for 30 min. Adsorb spill with paper toweling and remove to
autoclavable tray or bucket. Wipe spill area clean using 10% bleach. Wear gloves and use
forceps or dust pan and broom to pick up any large pieces of broken glass.
         B. In the event of injury or exposure, carefully and thoroughly wash the injured or
exposed area with clean water for up to 15 minutes. Notify the Principle Investigator and the
Biosafety Officer, Daniel Eisenman (2-4304, eisenman@musc.edu). Proceed to
employee/student health or the ER if after hours. Injuries should be reported to Employee Health
Services by submitting an ACCORD form via the following site:
http://mcintranet.musc.edu/ehs/work_related.htm

(Please print full name and sign below)

By signing below I attest that I have read and understood these safety instructions and agree to adhere to
these rules at all times. Furthermore, I feel I have been properly notified and trained of the hazards in this
laboratory.

Researcher _________________________________________                Date: ___________


Researcher _________________________________________                Date: ___________


Researcher _________________________________________                Date: ___________


Researcher _________________________________________                Date: ___________


Researcher _________________________________________                Date: ___________

								
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