Washington University in St. Louis, Environmental Health and Safety
Institutional Biological and Chemical Safety Committee/Office of Biological and Chemical Safety
Return completed form to:
Office of Biological and Chemical Safety
Environmental Health and Safety
Campus Box 8229
314-747-0309, FAX 362-6786; ehsibc@wusm.wustl.edu http://ehs.wustl.edu
Instructions for Completing Recombinant DNA and Hazardous Research Materials Protocol
PLEASE READ ALL INSTRUCTIONS! (Thank you!)
On behalf of the Institutional Biological and Chemical Safety Committee (IBC), the Office of Biological and
Chemical Safety (OBCS) requests submission of a biosafety protocol for research activities involving:
 Recombinant DNA molecules and recombinant DNA-containing organisms or cell cultures which are subject to the NIH Guidelines for Research Involving Recombinant DNA Molecules, including human gene therapy trials.*
 Pathogenic microorganisms or agents potentially infectious to humans and/or animals.
 Human blood, tissues, and cell lines as well as potentially infectious animal blood, tissues and cell lines.
 Carcinogens, mutagens, teratogens, drugs, and toxins when administered in vivo to animals or in vitro to cell or tissue cultures to induce a biological outcome.
 Animals for which a reasonable potential for transmission of zoonotic agents exists, e.g., wild-trapped animals, sheep, and rhesus macaques.
 Exotic plants, animals or microbes (e.g., non-indigenous plant or insect pathogen, or biological control agent); provide a copy of any required federal permit.
This document provides OBCS and the Institutional Biological and Chemical Safety Committee a detailed description of regulated or hazardous research elements and their management, providing a basis for risk assessment. A single biosafety protocol may be submitted to cover multiple related grant submissions. Once registered and assigned a safety committee registration number (IBC#), the protocol is valid for five years.
Modifications occurring during that 5-year period in the materials and methods and/or administrative changes, such as additional grant applications, staffing and location changes, should be submitted by including the changes on the original application form using a different font or color to indicate what has changed.
The RESEARCH ELEMENTS sections cover the use of recombinant DNA molecules and potentially biohazardous components of typical research projects. Skip sections that do not apply. Please complete the
DISPOSAL section for all protocol submissions. In RESEARCH PROTOCOL section, please provide an overview of the project and a detailed description of the practices employed in the management of biohazardous elements; discuss safety aspects of facility, containment equipment, personnel practices, and staff training that will ensure safe conduct of the investigation.
* The Recombinant DNA Transfer to Human Subjects Protocol is to be completed for projects involving transfer of recombinant DNA to human subjects. Please contact the Office of Biological and Chemical
Safety, Environmental Health and Safety, at 7-0309.
January 2011

References
Biosafety in Microbiological and Biomedical Laboratories. CDC/NIH. 5 th edition, February 2007. http://www.cdc.gov/biosafety/publications/bmbl5/index.htm
Bloodborne Pathogens: Exposure Control Plan. 1998. Washington University in St. Louis, Environmental Health and Safety. http://ehs.wustl.edu/resources/pages/default.aspx
Handbook of Environmental Health and Safety (“Blue Book”). 2000. Washington University in St. Louis,
Environmental Health and Safety. http://ehs.wustl.edu/resources/pages/default.aspx
Laboratory Standard. 1990. Department of Labor, Occupational Safety and Health Administration. 29 CFR, Part
1910.1450. Federal Register Vol. 55, No. 21.
NIH Guidelines for Research Involving Recombinant DNA Molecules. Revised January 2001 and subsequent amendments. National Institutes of Health. http://oba.od.nih.gov/rdna/nih_guidelines_oba.html
Occupational exposure to bloodborne pathogens; final rule. 1991. Department of Labor, Occupational Safety and
Health Administration. 29 CFR, Part 1910.1030. Federal Register 56(235).
Proposed Guidelines for Research Involving the Planned Introduction into the Environment of Organisms with Deliberately
Modified Hereditary Traits. 1991. USDA. Federal Register Vol. 56, No. 22.

Administrative Use Only
Registration Number Review Date
IBC#
I.
Washington University in St. Louis, Environmental Health and Safety
Institutional Biological and Chemical Safety Committee(IBC)/Division of Biological and Chemical
Safety(DBCS)
Return completed form to DBCS, Environmental Health and Safety Office, Campus Box 8229; fax to 362-6786; or e-mail to ehsibc@wusm.wustl.edu
CORE REGISTRATION INFORMATION
Name of Principal
Investigator (PI):
Job Title:
Office Phone:
Department:
Division:
Campus Box :
Email Address:
Lab Phone: Fax:
Name of Co-PI(s):
Contact Person: Email:
Protocol Type (check one) Applicable Registration Number & Review Date
New Amendment
General Protocol Title:
Renewal
Grant Title(s):
Signature of Principal Investigator Date
Amendment or Renewal IBC#
Granting Agency(s):
Chair, Institutional Biological and
Chemical Safety Committee/Biological
Safety Officer
Review date
Grant #:
Date
January 2011

II. RESEARCH FACILITIES
Location:
Where are experiments performed? Is the facility equipped with requisite equipment for containment and/or decontamination [e.g. autoclave, containment facilities or biological safety cabinets, (BSCs)]? (NOTE: The IBC requests prior notification, via written amendment to this protocol, regarding any change in location of containment labs, containment animal rooms, and/or BSCs.)
Building Name
Room number
Use of Room
(animal housing, lab, surgery, necropsy, growth chamber, greenhouse, other)
Containment
Equipment
(e.g. autoclave, biosafety cabinet, fume hood)
Equipment
Certification date(s)
III. LABORATORY PERSONNEL
List personnel involved with work covered under this research registration; include lab personnel: investigators, students, and research staff. If any of your staff is in need of annual training, online training can be found at http://ehs.wustl.edu/training/Pages/default.aspx
.
Last name/First name Job Title Phone number Date Annual EH&S
Lab Safety Training
Received
H EALTH S TATUS
Some unusual circumstances warrant special considerations or measures to prevent infection of laboratory personnel by certain microorganisms and other potentially etiological agents:
Please be informed (and ensure that all staff members are similarly informed) that certain medical conditions increase your risk of potential health problems when working with pathogenic microorganisms and/or animals. These conditions can include: pregnancy, immunosuppression, animal related allergies and chronic skin conditions. If any of these conditions applies, inform your personal physician/health care professional about your work.
January 2011 2

IV. RESEARCH ELEMENTS
Section IV.A: Research Description
Please provide a brief description or abstract of your research objectives and experimental design.
Section IV.B: Recombinant DNA
Table 1: Gene Source(s)
Under the “Risk Assessment” column, please indicate the perceived risk of the gene being expressed as low,
medium or high. Please refer to the following website when evaluating the risk of genes being expressed: http://atlasgeneticsoncology.org/Genes/Geneliste.html
. If medium or high is indicated, please provide an explanation for your assessment in the section under the table.
Gene/siRNA Name
(e.g. GFP - green
Gene Source(s)
(Genus, species, strain)
Function of Insert/Protein
Expressed or Silenced
Risk Assessment
(low, medium, or fluorescent protein) high)
Risk Assessment explanation: (e.g. tumor suppressor, oncogene, etc.)
Table 2: rDNA Use
Gene Name Sequencing/PCR only Plasmid
Table 3: Vector Description(s)
Vector Type
(plasmid, phage, etc.)
Vector Source
(e.g. vector lineage)
(Genus, species; if plasmid or viral)
Expression in:
Viral Vector Cell Culture
Technical Name of Vector
Provide reference or source
(if commercially available)
Animal/Plant
Risk Attenuation
(Replication defective?
Helper virus?
Disarmed?
K-12 derivative?
Reversion potential?)
January 2011 3

Table 4: Viral Vectors
Virus System
(Lentivirus, adenovirus, etc.)
Plasmids
Used
(List all)
Tropism 1
(e.g. pantropic, amphotropic, or ecotropic)
Transgenes expressed
(List)
Virus administered to:
Animals/Plants
(List)
Cell lines/tissues
(List)
1 Ecotropic – only infects murine or rat cells; amphotropic – infects human cells in addition to rodent cells; pantropic – infects all host species and cell types (e.g. VSV-G).
Table 5: Viral Vector Packaging (List cell lines used for packaging each viral vector.)
Virus System Packaging Line
Indicate where virus will be packaged and produced:
In Lab If this is checked, please describe the virus production procedures in Section V.C: Safety Narrative.
Other (please list):
Section IV.C: Microbiological Agents (including viral vectors)
Identify agents: Mark (y/n) in appropriate categories:
Microbe Source
(Genus, species, strain)
Human
Pathogen
Animal
Pathogen
Plant
Pathogen
Toxin
Production
Administered to animals in vivo
Recipient of rDNA
Large-Scale
Production
>10 liters
For bacteria which are human or animal pathogens, list any antibiotic resistance introduced (transiently or permanently):
January 2011 4

Section IV.D: Organ, Tissue, Cell Cultures, or Clinical Specimens (OTCC)
Cell line/Tissue
(Genus, species, strain)
Technical
Name
(e.g. NIH3T3,
Hep2)
Passage
(primary, established, immortal)
Administered to animals in vivo
(Yes/No)
Recipient of rDNA construct
(Yes/No)
Recipient of
Microbe
(Yes/No)
Recipient of
Chemical
(Yes/No)
For primary human cell lines and tissues, provide the source (e.g. clinic, Red Cross, etc.) and whether or not the cell lines/tissues are screened for any pathogens.
How screened? For which pathogens? Cell line/Tissue Source
Section IV.E: Hazardous Chemicals* or Biological Toxins
(* Toxins, carcinogens, mutagens, teratogens of proven or potential hazard, eliciting serious chronic or acute effects in humans, other animals or plants and requiring special handling precautions to prevent exposures.
Administered to:
Chemical Name
Nature of Chemical
(carcinogens, mutagens, teratogens, toxins, etc).
Administration
Route (IV, IM,
IP, SubQ etc.)
Highest Conc.
Administered
Maximum volume/ quantity handled
Microbe
(Yes/No)
OTCC
(Yes/No)
Organism
(Yes/No)
Section IV.F: Research Organisms - Vertebrates, Invertebrates, or Plants
Research Organism
(Genus, species, strain)
Used ONLY as source for cells/tissue
(Y/N) rDNA construct
(germ line or somatic?)
Recipient of:
Microbe
(Yes/No)
OTCC
(Yes/No)
If using transgenic animals, please list the source (e.g. WU mouse genetics core, generating in-house, collaborator, commercial supplier, etc.):
For administration of microbes to animals, please complete the following table:
Agent Route Dose
Chemical
(Yes/No)
January 2011 5

Section IV.G: Specialized Equipment
Please list any specialized equipment (e.g. FACS, imaging, injection apparatus, etc.) used in your research.
Equipment Location
V. RISK ASSESSMENT AND SAFETY PRECAUTIONS
Section V.A: Biological Risk Assessment
Project biosafety level (For descriptions, see http://www.cdc.gov/biosafety/publications/bmbl5/index.htm
)
BSL-1 BSL-2 BSL-2+ BSL-3
Personal Protective Equipment (PPE) – check all that apply
(Items below apply to work in the laboratory only, and do not include the animal facilities.)
DCM mandated PPE must be worn when working in the animal facilities.
Single gloves Double gloves Lab Coat Disposable lab coat/gown
Eye protection Respirator (e.g. N95 or PAPR) List type:
Avoidance of sharps Other (list):
Containment equipment
Biosafety cabinet used? Certification date
Yes No
Used during which activities?
List any other containment equipment used:
Section V.B: Chemical Risk Assessment
Personal Protective Equipment (PPE) – check all that apply
(Items below apply to work in the laboratory only, and do not include the animal facilities.)
Single gloves Double gloves Glove type: (latex, nitrile, etc.)
Eye protection Lab Coat
Respirator (e.g. N95) List type:
Other (list):
Containment equipment
Fume hood used? Certification date
Yes No
List any other containment equipment used:
Disposable lab coat/gown
Used during which activities?
January 2011 6

Section V.C: Safety Narrative
Because some situations require more description than can be captured using check boxes, you may need to provide additional information here. If you are using viral vectors, please discuss the potential adverse effects to personnel if they are exposed to the viral vector (taking into consideration both the vector and the transgene).
If you work with exempt amounts of select agent toxins, please discuss handling, storage, and inventory logs.
For hazardous chemicals, please provide a description of how you prepare and handle the chemicals (PPE, containment equipment, etc.). If you are using any specialized equipment, please describe how you will decontaminate the equipment following use (if applicable). Please use this space for any additional information requested by the IBC.
Section V.D: Disposal (For recommendations, see http://ehs.wustl.edu/committees/Pages/IBC.aspx#suggestedlanguage )
Please note that in accordance with the NIH Guidelines all recombinant DNA-containing material must be disposed of as biohazardous waste.
Disposal substance Disposal method
Solid biohazardous waste
Liquid biohazardous waste
Sharps
Chemical waste
Animal carcasses/bedding
Section V.E: Surface disinfection
Soap & Water 10% Bleach 70% Ethanol
Other (specify):
Section V.F: NIH Guidelines Assessment
List the relevant sections of the NIH Guidelines. The complete document is available at http://oba.od.nih.gov/rdna/nih_guidelines_oba.html
.
Section V.G: Exposure Prophylaxis
For each microorganism, consider the consequences of an accidental exposure (e.g., mucosal splash, inhalation, or inoculation), which might occur during experimental handling. Consider that organisms normally not pathogenic for healthy humans may become so when the natural barriers to infection are circumvented. Prepare a response procedure. It could be a simple matter of washing the wound with soap and water. If antibiotic treatment is recommended, please include information about the antibiotic of choice and its availability. The exposure response plan should be posted in the laboratory. Contact Employee Health at 314-362-3528 or, if after hours, the Infectious Disease Division 24-hour hotline at 314-747-3535.
January 2011 7

Viral Vector/Microbe Response
Section V.H: OSHA Bloodborne Pathogens Standard
Research involving the use of human-derived substances (e.g. blood or blood components, tissues, secretions) or human-derived cell lines (primary, established, or immortal), is subject to the OSHA Bloodborne Pathogens (BBP)
Standard. Have members of your lab been enrolled in the WUSTL Bloodborne Pathogens Program? If no, please contact WUSTL Environmental Health and Safety at 362-6816 or consult http://ehs.wustl.edu/RESOURCES/pages/default.aspx
. Please attach the completed “Verification of Training” page from Appendix 15 of the Exposure Control Plan, found at the web page above, to your submitted protocol or fax a copy to 362-1995.
YES : N/A:
Section V.I: Shedding of Hazardous Materials
If animals are receiving infectious agents or hazardous chemicals, please describe the source of any shedding or hazardous metabolites and any special safety precautions required (cage labeling post infection, additional PPE, etc.).
Microbe shedding/Hazardous chemical metabolites Special precautions (PPE, cage labeling, bedding changes)
Section V.J: Standard Operating Procedures
Chemical Hygiene Plan: Does your laboratory have a Chemical Hygiene Plan? If no, please contact WUSTL
Environmental Health and Safety at 362-6816 or consult http://ehs.wustl.edu/RESOURCES/pages/default.aspx
for assistance (follow link for Danforth or Medical
School “Blue Book”).
YES NO
Spill protocol: Will your laboratory be following the EH&S spill protocol (Appendix A)? If no, please append your spill protocol.
YES NO
Transport protocol: Will your laboratory be following the EH&S transport protocol (Appendix B)? If no, please append your transport protocol.
YES NO
VI. OTHER REGULATORY REQUIREMENTS
DOT HazMat Shipping Certification
The U.S. Department of Transportation requires that all persons involved in shipping hazardous materials in commerce be trained and certified in proper handling of these materials. The WUSTL Environmental Health and
Safety Program offers training to meet this requirement. Contact WUSTL EH&S
( http://ehs.wustl.edu/training/Pages/default.aspx
; 362-6816) to schedule training.
January 2011 8

Appendix A: Standard Spill Protocol
LARGE BIOHAZARDOUS SPILL
OUTSIDE A BIOLOGICAL SAFETY CABINET

Alert co-workers and leave lab area immediately.
 Close door, post lab with “
Do Not Enter
” sign.

Remove and put contaminated garments into a container for autoclaving.

Wash hands and face with soap or antimicrobial agent.

Notify supervisor.

Wait at least 30 minutes before re-entry (to allow aerosols to dissipate).

Upon re-entry, wear PPE (disposable gown, mask, gloves, etc.)

Carefully pour an appropriate disinfectant solution (1:10 dilution of household bleach) around spill and cover spill with disinfectant soaked paper towels. Let stand for at least 20 minutes.

Transfer all contaminated materials into an autoclave bag.

Wash and mop entire area with appropriate disinfectant

Remove & discard PPE. Autoclave.

Shower or wash hands with soap.

If the spill involves recombinant DNA, notify the Biological Safety Officer (747-
0309)
January 2011 9

SMALL BIOHAZARDOUS SPILL
OUTSIDE A BIOLOGICAL SAFETY CABINET

Cover spill with paper towels and carefully pour an appropriate disinfectant solution
(1:10 dilution of household bleach) around spill, taking care to not create aerosols while pouring.

Cover spill with disinfectant soaked paper towels. The operator should be wearing gloves during this procedure.

Let stand for at least 20 minutes.

Transfer all contaminated materials into an autoclave bag.

Wash hands with soap or antimicrobial agent.

Autoclave all contaminated materials.
January 2011 10

BIOHAZARDOUS SPILL
INSIDE A BIOLOGICAL SAFETY CABINET

Immediately stop all work. Leave BSC blower fan on during clean-up.

Cover spill with paper towels and carefully pour an appropriate disinfectant solution around spill. Cover spill with disinfectant soaked paper towels. The operator should be wearing gloves during this procedure.

With paper towels & appropriate disinfectant detergent, wipe down BSC walls, work surfaces, and equipment.

Flood work surface & drain pan (Type II BSC) with disinfectant. Allow to stand at least 20 minutes.

Wipe up all excess disinfectant.

Autoclave all contaminated materials
January 2011 11

Appendix B: Standard Transport Protocol
T RANSPORTATION OF R ESEARCH AND C LINICAL S PECIMENS
 Research/clinical specimens should be transported in a sealed, leak-proof primary container within a sealed, leak-proof secondary container (e.g. Tupperware container).
 The secondary container should be labeled with the universal biohazard sticker or carcinogen sticker as appropriate (available from the EH&S office).
 The outside of the secondary container should be disinfected (for biohazards) or cleaned (for chemical hazards) so that the use of personal protective equipment is not required.
 Public areas and elevators should be avoided if at all possible. If public areas/elevators must be used, gloves must not be worn. If necessary, the hand holding the specimen may be gloved, but under no circumstances should gloves be used on common surfaces such as elevator buttons.
 Even though a spill is unlikely when this protocol is followed, a spill protocol should be prepared in advance. The area of the spill should be isolated and EH&S should be contacted (2-6816; after hours, 2-
HELP) to clean up the spill.
January 2011 12