Institutional Biosafety Committee
Application for Approval of Research or Instructional Activities Involving Recombinant DNA or
Other Biohazardous Materials
This application should be submitted to the Butler University Institutional Biosafety Committee c/o the
Institute for Research and Scholarship, Jordan Hall 109D, 317-940-6424.
Primary Investigator
E-mail address
Phone number
Office
Department/College
Title of Research
In what room(s) do you plan to work with, store or dispose of recombinant DNA? (Example: GH 48 sterile transfer of recombinant bacteria; GH 40 - cloning procedures; GH 10 - autoclaving of wastes; GH
42 - storage of recombinant cell lines)
What vector(s) do you plan to use? Give the class, vector name, and vendor or other source. (Examples:
plasmid pSE280, Invitrogen, lambda phage, Ziplox, Promega)
What gene(s) do you plan to use? Be sure to include the source organism. (Examples: mouse
androgen binding protein, cDNA library from spinach roots, Chlamydomonas reinhardii, small subunit of
ribulose-1,5-bis-phosphate carboxylase/oxygenase)
What host organism(s) do you plan to use? (Examples: Escherichia coli Y 1089 (r-); armyworm cell
culture; Arabidopsis thaliana plants.)
Project description
Attach a brief description of how you plan to use the recombinant genes. State whether this activity will be
used in an instructional manner (for a class) and/or for research purposes. Also include a brief
description of how accidents involving the rDNA material and/or vector host organisms will be handled
(such as applying bleach to the benchtop area of a small spill)
(Example description: In order to study subcellular targeting of pyruvate decarboxylase PDC2 in tobacco,
I will make a fusion between the cauliflower mosaic virus 35S promoter, the Arabidopsis thaliana pyruvate
decarboxylase PDC2 coding sequence, and a blue-shifted green fluorescent protein gene. Initial cloning
steps will be carried out in vector pBluescript using host Escherichia coli XL1-Blue (Stratagene), and
selection on ampicillin. The gene fusion will be pasted into shuttle vector pGA748, and this construct will
be electroporated into Agrobacterium tumefaciens LBA4404. Agrobacterium clones selected on
kanamycin will be used for tobacco leaf disk transformation. The distribution of green fluorescent protein
in transformed tobacco callus cells will be examined by fluorescence microscopy. Any spillage containing
E. coli or Agrobacteria will be soaked into paper towels, which will be added to other accumulated
biohazardous waste in labeled red bags and decontaminated via autoclaving; the surface(s) exposed to
the spill will be treated with bleach.)
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Revised 9/2015
Institutional Biosafety Committee
Application for Approval of Research or Instructional Activities Involving Recombinant DNA or
Other Biohazardous Materials
Safety assessment and signatures
Safety Precautions for Laboratories using recombinant DNA (rDNA)
Laboratory Access
Only authorized persons may enter the laboratory. An authorized person must understand the safety
precautions of a recombinant DNA laboratory or be accompanied by one who does. Laboratory doors
should be kept closed and locked except when a laboratory worker is present. All entrances to the
laboratory from the hallway should be marked with contact information regarding the faculty member(s) in
charge for the use of emergency personnel
Protective clothing
Goggles or safety glasses should be worn in the laboratory by all personnel at all times.
Closed Toe Footwear should be worn in the laboratory by all personnel at all times. No sandals, flipflops, etc.
Laboratory coats should be worn in the laboratory whenever rDNA work is being done. Do not wear
these coats into areas in which food or drink may be consumed. Isolate them in a plastic bag when
taking them to the laundry.
Disposable gloves must be worn when handling microorganisms or nucleic acids. Change gloves
immediately after obvious contamination or tears. Do not leave the laboratory and handle door-knobs,
etc. while still wearing gloves. Wash your hands before leaving.
Goggles or safety glasses with a UV-protective coating are required whenever gels are observed on
an ultraviolet transilluminator. Both goggles AND a face shield are required for longer exposures, such as
cutting out bands. Eye damage and serious facial “sunburn” may result if these precautions are ignored.
Masks should be worn when weighing out or cleaning up dangerous powders such as ethidium bromide,
acrylamide and SDS. Always read labels before handling chemicals.
Laboratory behavior
Food, drink, gum-chewing, smoking, and application of cosmetics is prohibited in the laboratory.
No food or drink may be stored in refrigerators or elsewhere in the laboratory.
Mouth pipetting is prohibited.
Work Surfaces and Spills
Bench surfaces and microcentrifuges must be disinfected with a suitable agent (such as 70% ethanol,
Lysol, or bleach) after use and after any spill of viable material. An absorbent lab mat may be used on
areas in which spills are possible; place any contaminated lab mat in the autoclave buckets.
Spills of microorganisms should be cleaned by absorption into paper towels followed by disinfection of
the surface with a suitable agent. Place all soiled paper towels and gloves used during spill clean-up into
autoclave buckets. If a spill is too large for simple clean-up, such as a broken culture flask, leave a
warning sign and contact the laboratory supervisor.
Contaminated clothing should be removed and replaced with emergency scrubs kept in the laboratory.
Use the bag from the scrubs to store clothing until it can be laundered (with bleach) or disposed.
If material gets on skin, wash skin thoroughly with soap and water. If material gets in the eyes, rinse for
fifteen minutes with saline solution or eye wash. Contact laboratory supervisor.
Waste disposal
Solid wastes contaminated by microorganisms or rDNA, including used gloves, pipette tips, Petri dishes,
and paper products, must be placed in an autoclave bucket lined with paper towels or in an autoclavable
biohazard waste bag. These wastes should be autoclaved at the P6 setting. Autoclavable bags should
be placed in a bucket or tray for autoclaving. After autoclaving, solid wastes should be transferred to the
green bin in GH10 for disposal. The bin is marked “Autoclaved Solid Wastes”.
Liquid cultures in containers less than 250 ml/container should be decontaminated by autoclaving at P5
or P6. Liquid cultures in containers 250 ml and up must be autoclaved at P6. Alternatively, small
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Institutional Biosafety Committee
Application for Approval of Research or Instructional Activities Involving Recombinant DNA or
Other Biohazardous Materials
quantities of liquid culture may be decontaminated by addition of one volume of bleach followed by
soaking overnight. Decontaminated liquid waste may be washed down the drain.
Sharps such as scalpel blades and glass waste must be placed in a sharps container.
Chemical wastes must be placed in appropriate waste containers. Liquid wastes should be segregated
as either aqueous, halogenic organic (like chloroform), or flammable organic waste. Solid wastes should
be segregated as hazardous or non-hazardous. Gels containing ethidium bromide or similar mutagens
should be placed in a labeled plastic bag and allowed to dry. Buffers containing ethidium bromide or
similar mutagens should be passed through a sealed charcoal filter. Used filters and dried gels can be
submitted to the Environmental Programs Office (X6408) as hazardous wastes.
Safety:
I have read and understood the precautions detailed in the Butler University policy on
“Safety Precautions for Laboratories Using Recombinant DNA.”
This project requires no precautions beyond those detailed in the Butler University policy
on “Safety precautions for laboratories using recombinant DNA.” I have read,
understood, and will follow the guidelines in the policy document.
In order to minimize health and/or environmental risk factors specific to this project, I will
take precautions in addition to those in the Butler University policy document. A
discussion of the project-specific risks and precautions is attached.
This project does not require all of the precautions in the policy document. An explanation
of any adjustments to the standard precautions is attached.
Student Involvement:
I will be supervising student research on this project. I will train any student involved in
this work such that they understand how to comply with the Butler University policy on
“Safety precautions for laboratories using recombinant DNA.” Each student working on
this project will submit a signed (by both the student and myself) "Institutional Biosafety
Committee's Qualifications Form for Student Investigators" form to the University
Research Programs Office
I will NOT be supervising student research on this project.
Maintaining Compliance:
I will report to the IBC any change in personnel (particularly students) working on this
project.
I will report to the IBC any change in the class of vector used. (such as switching from a
plasmid vector to a viral vector)
I will report to the IBC any change in the source organism(s) used in this project.
I will report to the IBC any change in the host organism(s) used in this project.
Signature of Primary Investigator
Date
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Revised 9/2015
Institutional Biosafety Committee
Application for Approval of Research or Instructional Activities Involving Recombinant DNA or
Other Biohazardous Materials
IBC Action:
□
Project approved as presented.
□
Project approved with modifications.
□
Additional information requested.
□
Approval denied.
Signature of BIRS Director_
Date
Signature of IBC Chairperson_
Date
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