ISSUE
FNR
Hazard Committee
PFEN 158
Ju l y
2009
Orientation
Communication
Lab Safety
topics
Your Guide to
Lab Safety
Safety in this Laboratory P.1
Waste Collection P.2
This pamphlet will serve as
Laboratory Procedures P.3
your introduction to and
Personal Protective Equip. P.4
reference for safety concerns
while working in this
Further Information P.6
laboratory. Please read and
become familiar with this
information. Most general
guidelines and safety issues
have been addressed, but
there are always new
situations that may arise.
This guide is not a substitute
for proper training in any
unfamiliar areas. Always seek
assistance when warranted.
All procedures and guidelines,
as laid out here, must be
followed irrespective of your
previous laboratory
experiences or personal
preferences. It is acceptable
to take additional precautions
if desired but do not use
alternate procedures without
prior approval of the lab
manager or the faculty
member in charge.
Safety in this Laboratory
What follows is a general overview of the
safety guidelines established for the
Molecular Tree Physiology Lab in Pfendler
Hall, room 158. If you have any questions,
do not hesitate to ask the lab manager (43569).
 Lab Attire: Lab coat, gloves, eye protection
as appropriate. Shorts OK. NO open-toed
shoes, sandals or flip-flops.
 Food: No food or drinks allowed in the lab.
There is a table outside the door to store
food/drinks.
 Safety Manuals: Purdue Radiological and
Environmental Management (REM) safety
handbooks must be read/understood and a
form signed and returned indicating such.
 Sharps: Broken glass and plastics
(ANYTHING that could puncture a plastic
bag) must be discarded into a cardboard or
plastic container. Metals (razor blades,
needles, etc.) must be discarded into plastic
containers. All containers must be labeled
clearly as SHARPS. Any chemically or
biologically contaminated sharps must be
contained separately and labeled as such.
 Open Containers: No open containers
holding any reagents are allowed. All
containers must be closed/covered in a way
to prevent spillage.
 Labeling: All containers (chemical, waste or
otherwise) must be labeled with the contents.
All containers you generate should be labeled
with at least your name, the date and the
1
contents.
 High Liquids: REM stipulates that all liquids
in containers stored above your eye level
must be obtained with the use of a step-stool.
Please be aware of this and store liquids
lower than eye level if possible.
 Dishes/Sinks: Please either wash your
dishes as you make them or collect them in
the area designated for the undergrads to
wash. Do not leave dishes in the sinks.
Clean up the sink if you have a mess. Do not
leave chemical residues, gel fragments, tape,
parafilm, tips, broken glass, plant matter,
paper towels or anything else in the sinks.
 Contamination: Any samples (plates, boxes,
cultures, etc.) that are contaminated should
be sterilized and removed from the lab as
soon as possible.
 Biohazard Waste: Collect all biohazard
waste into the designated containers. The
waste will be autoclaved, packed into plastic
bags and boxed for disposal by REM. Feel
free to collect and autoclave waste when a
container is full.
 Hazardous Waste: All chemically
contaminated waste (solid and liquid) should
be collected in labeled containers. Waste
may generally be combined as long as it is
fully labeled. REM must be contacted for
waste removal.
 Ethidium Bromide: Ethidium bromide (EtBr)
may be used for staining electrophoresis DNA
and RNA gels. EtBr must never be mixed
See Safety, page 2…
Injuries
All significant injuries should be
reported immediately to the lab
manager or the FNR business
office in PFEN 126. Minor
injuries which can be
satisfactorily cleaned, disinfected
and bandaged using the lab firstaid kit (under the center sink) are
not required to be reported.
Injuries such as deep cuts,
punctures, burns, falls, etc.
should be reported. If there is
risk of infection (due to bacterial
Safety… continued
into molten gels or gel buffer
for use during
electrophoresis. Gels must
be soaked in a reusable
EtBr solution kept in a
dedicated and labeled
container. Residual EtBr in
the rinsate from empty
containers may be disposed
down the sink drain. Never
touch or operate any
equipment (computers,
cameras, faucets, drawers,
etc.) while wearing gloves
contaminated with EtBr.
Other people will be using
these items without gloves.
 Gel Boxes: Clean and rinse
all electrophoresis gel boxes
and related equipment when
finished. Do not leave a
contact or otherwise), then it
 Know the locations of the
safety showers, eye washes
and fire extinguishers in your
work area. If you are visiting
an unfamiliar lab, determine
these locations prior to
working.
In addition to the items
mentioned so far, you
should make note of and
adhere to the following as
well:
 Never ignite alcohol on any
lab work surface, especially
within fume hoods, sterile
hoods or biological safety
cabinets. Free liquids
cannot be controlled and the
flames are often sucked into
the ventilation systems,
posing a fire hazard.
 Always wear appropriate
safety garb (lab coats,
goggles, gloves, etc.).
should always be reported.
 Do not attempt to use
unfamiliar equipment. Always
request training for something
with which you are not
thoroughly familiar.
 Keep the lab tidy. Clean up
spills, properly dispose of
trash and used materials,
clean up equipment when you
have finished with it and put
items back where they came
from.
 Use common sense when
completing any task. Do not
attempt to rush any
processes or equipment.
Allow enough time to
complete what you need to
do properly.
Reporting is for your own
protection, should complications
arise at a later date. It is also
University Policy.
If an injury is severe or occurs
after hours, obtain whatever
medical attention is warranted
and report the injury to the
manager or office as soon as
practical.
Your supervising faculty should
be informed of the injury in a
timely manner.
For all Emergencies
 Call 911 for emergency care.
 The Student Health Center is
not for work related injuries.
 Use Clarian Arnett Occ. Health
Center (448-8708) or Regional
Occupational Care Center
(446-2450) for general care.
 Use Clarian Arnett Hospital,
Home Hospital or St Elizabeth
Medical Center only for
emergencies.
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mess for the next user to
clean-up. Do not store gel
buffers in the gel boxes. If
they will be reused, they
must be stored in tightly
closed, labeled containers.
 Fume Hoods: Do not
overcrowd the fume hoods.
This will restrict air flow.
Chemical waste containers
may be stored in the fume
hoods.
 Lab Doors: Laboratory
doors should be kept closed
for proper fume hood
operation. Keep the doors
locked whenever the lab is
unoccupied. Do not leave
valuables in the lab.
 MSDS: Material Data and
Safety Sheets are available
in the lab and in the file
cabinet in the Pfendler
loading dock (next to G058).
Waste Collection
All bio-hazardous and
chemically hazardous waste
generated in the laboratory is
collected and disposed of by
Purdue’s Radiological and
Environmental Management
service (REM). Prior to
collection by REM, the
materials generated must be
properly contained, labeled
and/or decontaminated/
sterilized.
Bio-hazardous waste must be
placed into designated waste
containers (separate
containers for sharps versus
solid waste). All waste should
be autoclaved prior to
disposal. If liquid or potentially
liquid waste is autoclaved, it
should be contained within a
tote to collect any spillage of
liquid.
After autoclaving, the bags of
waste must be packaged into
a cardboard box lined with a
plastic bag (both provided by
REM) and sealed with tape. A
biohazard waste removal form
should be filled out and
attached to the box and REM
contacted for removal. Sharps
boxes, once closed, may be
autoclaved and placed in the
same box as the bags.
Chemically hazardous waste
should be contained in an
appropriate container for its
state (solid or liquid). All
containers must be labeled with
all waste contained within.
Containers should be tightly
closable and unlikely to spill
when removed. REM should
be contacted for removal prior
to accumulating large amounts
of waste or completely filling
bottles. Chemically hazardous
waste may be stored in waste
containers in the fume hoods.
See Waste, page 3…
Proper Waste
Storage
All waste generated by a
laboratory is to remain in that
room until it is picked up for
proper disposal by REM.
Quantities of collected waste
should be minimized (such as
Waste… continued
Bacterial cultures may be
decontaminated by incubating
them at room temperature for
thirty minutes with an equal
volume of a 10% bleach
solution. The resulting
suspension may then be
flushed down the sink drain.
DO NOT bleach soak pipette
tips or tooth picks. Put them
directly into the red sharps
containers.
Any bulk amounts of agar
waste ([plant] plates, culture
boxes, culture tubes, etc.)
should be autoclaved as a
liquid then contained in a
double layered plastic bag and
allowed to cool/solidify before
removal as waste.
Non-contaminated sharps may
be boxed, sealed, labeled
clearly as SHARPS and
placed beside the dumpster.
Laboratory Procedures
These procedures should be
followed when working in this
laboratory. Please feel free to
ask before doing something if
you are unsure.
 Generally, you should
prepare and maintain your
own reagent, media, tube, tip
and culture box stocks. If
you are in need of something
and wish to borrow material
from another lab member,
always ask for their
permission first. Never
assume that you may
perpetually use someone’s
supplies.
 Most people will expect
that any sterile materials
(common or borrowed) will
be opened and used in a
sterile environment, such as
the sterile hood. Unless you
have been explicitly informed
otherwise, you should
nothing over 20 gallons at one
location).
maintain these conditions.
 Regularly check your
culture containers and
stocks for contamination.
 All contaminated plant
culture materials (plates,
boxes, tubes, etc.; with or
without plants) should be
removed from the general
population as soon as
possible. They should be
stored away from other
materials until they can be
autoclaved. Currently, they
are kept under the counter,
across from the ultra-low
freezer.
 Never open contaminated
cultures in the lab,
especially in the sterile
hoods. Contaminated
materials need to be
autoclaved prior to opening
them for disposal.
Chemical waste should be
segregated by chemical type
and reactivity. Containers
should be generally labeled for
Acids, Bases, Organics and
Aqueous wastes. This is in
addition to the specific labeling
of the waste constituents.
Biological waste should be
stored in an orderly contained
manner.
See Procedures, page 4…
Paperwork
The following certification forms and
All informational booklets and guides are available on the
training sessions must be completed by
Purdue Radiological & Environmental Management (REM)
anyone working in this lab. They should be
website at:
completed as soon as possible.
http://www.purdue.edu/rem/home/files/guide.htm
They should not be stored
haphazardly…
Specifically, the Biological Safety Guide is at:
 Biological Safety Awareness
 Chemical Hygiene Awareness
 Biological Exposure Occupational Health
 Personal Protective Equipment Awareness
 Pesticide Exposure Training
http://www.purdue.edu/rem/home/booklets/bioman.pdf
The Chemical Hygiene Guide is at:
http://www.purdue.edu/rem/home/booklets/CHP2003.pdf
Information regarding Pesticide Exposure Training is at:
http://www.purdue.edu/rem/ih/wps.htm
Sharps should be collected in
appropriate containers…
 Meeting with Lab Manager
3
Pesticide
Exposure
Training
All university employees and
students that spend any time in
the greenhouses or in an
environment that may potentially
bring them into contact with
pesticide or herbicide use are
required to attend an educational
training seminar discussing the
specific hazards present and
proper precautions to be taken.
Training required only once
every five years. See For
Further Information on the last
page to schedule a session for
yourself or one of your
employees.
Biological
Exposure
Occupational
Health Program
All university employees and
students working in an
environment which may bring
them into contact with a
biohazard have the option of
participating in this program. It
provides specific information and
screenings for the particular
hazards you may encounter.
Participation is optional, however
submission of the participation/
declination form is required.
Procedures...
continued
 Do not leave any of the
ultraviolet sterilization lights
on while people are in the
lab. They should only be
used overnight, when the lab
is empty. The last person
out of the room at night
should turn on the UV
light(s) and the first person
in should turn them back off.
The sash should be lowered
completely when the UV
light is on and raised when it
is off.
 There are signup sheets
on some pieces of the more
heavily used lab equipment.
Please use them to reserve
a time slot and/or to let
others know who to contact
if there is a problem.
 Never put liquid nitrogen
into the sinks. Avoid putting
dry ice into the sinks as well.
The extreme cold can cause
the basins to crack.
 There are autoclaves
available in room 141, 258
and the WSLR subbasement. There are signup
sheets for the 141 unit.
Please make arrangements
with the lab in 258 prior to
use. The WSLR units are
public access.
 As a courtesy, always ask
the lab manager of room
141 prior to using any of the
equipment in that lab.
 Try not to let the ultra-low
freezer stand open for
extended periods of time. If
you need to search in a
container, remove it and
close the door first.
 Liquid nitrogen dewars
may be filled by taking them
to the Pfendler loading dock
and filling out the request
form. Contact the lab
manager for an account
number
 If you need to order
materials, please contact the
lab manager with your
request, preferably via email.
 Any other questions,
please ask.


4
Personal Protective
Equipment
Personal protective
equipment (PPE) is provided
in the lab and should be used
as needed to prevent possible
injury. The following
describes the equipment
relevant to activities in this
lab and provides basic usage
information. If you are not
already, you should become
familiar with the proper use
of these items.
If at any time you have
questions regarding the need
for or proper usage of
protective equipment, please
contact the lab manager.
hand, then remove
remaining glove by
reaching under cuff and
grasping the
uncontaminated surface.
 Cryogenic Gloves: Gloves
should be worn anytime you
may come in contact extreme
cold (dry ice, liquid nitrogen,
etc.)
o These gloves are not for
immersion.
o Donning/Doffing:
Inspect for holes or tears.
Do not use if they do not
fit well. Use care to avoid
 Chemical/Biological
Gloves: Gloves should be
worn anytime you may come
in contact with potentially
undesirable or hazardous
substances.
o You generally should
wear gloves for virtually
every procedure to do in
the lab. You should not
wear gloves while using
common devices, such
as the telephone and
the computers. You
should not wear gloves
when leaving the lab.
Take some with you if
you will need them.
o Donning/Doffing: Put
on and check for holes
or other flaws. Very
tight fits are undesirable.
Remove before leaving
or touching
inappropriate items. If
possible, clean prior to
removal. Remove one
glove with other gloved
freezing temperatures
when removing.
 Dust Mask: Dust masks are
merely for comfort when
dealing with non-hazardous
dusts and mists.
o These do not provide
protection against any
hazardous materials.
o Donning/Doffing: Apply
mask so that it fits snugly
over nose and mouth
prior to entering area.
See PPE, page 5…
PPE...
continued
Remove after vacating
area to avoid breathing
nuisance materials.
 Apron: Aprons protect the
user from chemical splashes
and nuisance exposure to
other potentially soiling
materials.
o Use an apron suitable
for the substance in
question. Corrosives or
organics may require a
more substantial
material than what is
used for washing
glassware.
o Donning/Doffing:
Inspect the apron for
holes or tears. Do not
use if the straps cannot
be secured properly.
Take care when
removing to avoid
contact with residual
chemicals. Clean after
using.
 Face Shield: Provides
splash, impact or radiation
protection.
o Use the proper type.
One splash shield will
not handle all situations.
If used for UV radiation,
make certain it actually
is rated for UV filtering.
o Donning/Doffing:
Inspect for cracks,
scratches or excessive
wear that might impair
function while using.
Make certain all areas to
be protected are
covered. Remove while
avoiding contact with
and potential
contaminants.
 Impact Goggles: Provides
eye protection from potential
flying debris.
o These are not for splash
protection.
o Donning/Doffing: Apply
with or without corrective
eyewear. Inspect for
cracks, scratches and
other wear that might
impair function while
using. Put in place
before starting
procedures.
 Splash Goggles: Provides
eye protection from chemical
spills and splashes. Use
when within ten feet of 50 ml
or more of a hazardous
liquid.
o These do not necessarily
provide adequate impact
protection.
o Donning/Doffing: Apply
with or without corrective
eyewear. Inspect for
cracks, scratches and
other wear that might
impair function while
using. Put in place
before starting
procedures. They
should fit snuggly
against face, with no
gaps around edges.
Clean after use.
should cover the leg to the
knee. Other protective gear
is generally necessary in
addition to regular clothing.
o Any clothing that is
contaminated with
hazardous materials
should not be taken
home for laundering.
Protective Gear
Some examples of PPE, in the
order described…
o Donning/Doffing:
Clothes should be worn
appropriately, buttoned
and with sleeves down.
Feet should be covered
by shoes at all time.
Sandals, flip flips and
the like are not allowed.
 Lab Coat: Protects against
harmful liquid splashes and
hazardous solid chemical
contact with skin or regular
clothes.
o These need to be
buttoned completely to
be effective. Sleeves
should be down.
o Donning/Doffing: Wear
as you would expect.
Avoid excessive liquid or
chemical contact. Do
not wear while soiled.
All areas to be protected
should be covered (to
knees preferably).
Remove with avoiding
contamination. Wash as
needed. Contaminated
coats should not be
laundered at home.
 Oven Mitt/Autoclave
Glove: Protects against
burns from hot surfaces.
o Does not protect against
flames or superheated
material. Provides
minimal protection from
steam and liquid
splashes.
 Donning/Doffing: Inspect
for excessive wear, holes
and tears. Do not use while
wet, as moisture will conduct
heat. Do not put on over
other protective gloves.
 Clothes and Shoes:
Clothing appropriate for the
task to be completed should
be worn. Feet should always
be covered. Shorts, if worn,
5
For Further Information
For additional safety related information, please see one of the following:
 Lab Manager:
Nathan Hilliard
PFEN G076
4-3569
nhilliar@purdue.edu
 Lab Faculty:
Rick Meilan
PFEN G021E
6-2287
rmeilan@purdue.edu
 MTPL Web Page:
http://web.ics.purdue.edu/~nhilliar
 Genetics Lab Manager:
Marcia Kremer
6-7685
marciakremer@purdue.edu
 REM Website:
http://www.purdue.edu/rem/
 Bio-Waste Disposal:
http://www.purdue.edu/rem/eh/biowaste.htm#disp
PFEN 141
o Pickup Request:
4-0121
o Request Form:
http://www.purdue.edu/rem/home/files/forms.htm#BMPU
 Chemical Waste Disposal Questions:
o Request Forms:
4-0121
http://www.purdue.edu/rem/home/files/forms.htm#HMM001
 Pesticide Training:
o Brian McDonald
6-3712
o Judah Young
4-7293
Molecular Tree Physiology
Laboratory
Purdue University
Pfendler Hall, Room 158
715 W. State St.
W. Lafayette, IN 47907-1061
Phone: (765) 494-3569
Fax: (765) 494-9461
E-mail: nhilliar@purdue.edu
Lab Safety
6
Issue
PFEN 158
Ju l y 2 0 0 9
Acknowledgment of Training
Training Progress
Please initial next to the
After review of the material provided to you, any
guidebooks referred to you and completion of any
needed additional training, you do here certify the
following has been completed and understood:
appropriate line to indicate your
completion of that step. If the
step does not apply, write NA on
Initial
the line. Be certain to ask for
additional training or
information if needed.
When all items have been
completed, sign and date the
form and return to the Lab
Manager.
You may write comments or
suggestions in the lower area.
Reviewed Lab Safety Information Packet Provided
____
Submitted Biological Safety Awareness Form
____
Submitted Chemical Hygiene Awareness Form
____
Submitted Biological Exposure Health Form
____
Submitted PPE Awareness Form
____
Obtained Additional PPE Instruction (if required)
____
Completed Pesticide Training Program (if required)
____
________________________________________
Print Name
Molecular Tree Physiology
Laboratory
Purdue University
________________________________________
_____
Signature
Date
Pfendler Hall, Room 158
715 W. State St.
You may write comments in this area:
W. Lafayette, IN 47907-1061
Phone: (765) 494-3569
Fax: (765) 494-9461
E-mail: nhilliar@purdue.edu
Lab Safety
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Issue
PFEN 158
Ju l y 2 0 0 9