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SAFETY TRAINING DOCUMENT FOR THE JUDELSON
LABORATORY
A. INTRODUCTION
At the University of California, a safe working environment results from cooperation between
the University, the faculty member, and the individual worker. This document will inform
you of the hazards associated with working in the laboratory, and provide information on
safe working practices and guidance in resolving safety concerns. If after reading this
document you are unsure of safe procedures for performing your work, request instruction
from Howard Judelson or the UCR Department of Environmental Health and Safety (8275528).
B. PROCEDURES FOR EMERGENCIES AND ACCIDENTS
1. GENERAL INSTRUCTIONS
In case of any emergency, FIRST determine if the incident poses an immediate danger to you. If
so, leave the area immediately. SECOND, determine if the incident presents an immediate
danger to those nearby. If so, evacuate that area. THEN, once you are in a safe area, call for
help by dialing 911.
If you need to evacuate the building, do so by activating a fire alarm. Don't take the elevator!
Meet outside the building. Someone may be there with a clipboard, to take your name.
2. PROCEDURES FOR MEDICAL EMERGENCIES AND ACCIDENTS
For serious and life-threatening accidents, call 911 for an ambulance. Otherwise, seek
medical attention at your own discretion, using the laboratory first aid kit or an outside
physician, or other entity. Please notify Howard Judelson of all accidents.
3. PROCEDURES IN CASE OF FIRE OR EXPLOSIONS
Leave the area of immediate danger, being sure that others are also out of the area. Close the
doors to the room involved. Activate the nearest building fire alarm to evacuate others from
the building. Call the Fire Department: Dial 911 (for your safety leave the fire zone before
calling!). Leave the building.
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Fighting fires: Do not attempt to extinguish the fire if it is large or if hazardous materials are
involved! You can attempt to extinguish fires if you are confident that you will be able to
put it out.
4. PROCEDURES IN CASE OF EARTHQUAKE
Never exit a building during an earthquake!
Take cover under a desk or table or in a strong door frame until the shaking stops.
When shaking stops, secure your area (turn off gas burners and gas cylinders, cap dangerous
chemicals) and then exit the lab, closing the door behind you.
Evacuate the building.
Do not re-enter the building until official clearance is given.
5. PROCEDURES FOR A CHEMICAL SPILL (See "Chemical Hygiene," Section, D2e,
below).
C. GUIDELINES FOR INJURY AND ILLNESS PREVENTION
1. GENERAL WORK GUIDELINES
Understand the procedures involved in your work before you begin. This means know how
to safely use the materials, chemicals, equipment, etc. in your work. IF YOU DON'T
KNOW, ASK!!!
UC policy requires that individuals must wear long pants, or the equivalent, at all times when in
the laboratory. You must also wear closed toe/heel shoes in the laboratory, i.e. no clogs or
sandals.
Long hair, clothing, or jewelry should be tied back or otherwise confined.
Know the location and how to use exits, safety showers, eyewashes, first-aid kits, fire
extinguishers, spill kits, fire blankets, alarm pull boxes, and telephones.
Follow work practices that reduce the possibility of muscle injury. Take frequent breaks from
repetitive activities! Never move anything that is too heavy for you to handle alone: ask
someone to help you! When lifting, get a firm grasp while keeping your back close to
vertical, and then lift with the legs, not with the back! When working at a computer, sit erect
with the screen and keyboard directly in front of you; muscle strain will be reduced if your
forearms and legs are kept parallel to the floor.
Follow good housekeeping practices. Clean up as you go and keep work areas, aisles, corridors,
and exits uncluttered. Maintain accessibility to eyewash/emergency showers, fire
extinguishers, and electrical panels.
Report all accidents, injuries, or unsafe conditions immediately to Howard Judelson, the
Department Chairman, or EH&S (reporting can be done anonymously if desired). Do not
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ignore an unsafe condition!! Solve the problem yourself or bring it to the attention of
Howard Judelson or EH&S!!
2. LABORATORY-SPECIFIC GUIDELINES
Use the proper safety equipment for your procedure. This could include a fume hood, biosafety
cabinet, shields, or other equipment.
Wear the appropriate personal protective gear where laboratory or experimental conditions
dictate. This includes lab aprons, lab coats, gloves, goggles, face shields, dust masks,
respirators, and other equipment. Lab coats and eye protection are required when you, or
people adjacent to you, are handing hazardous chemicals.
Wear eye protection. Safety glasses are required when working with chemicals. The wearing of
contact lens are not advised for laboratory workers!
Wear clothes that protect the body against spills, dropped objects, and other accidental chemical
contact. Thus open shoes, sandals, and bare feet are prohibited. If you wear shorts, cover
your legs with a long lab coat when working with radioactive materials or dangerous
chemicals.
NEVER NEVER NEVER pipette by mouth. Use mechanical pipetting devices or bulbs.
Working when tired or very late at night is not advised. Fatigued workers cause accidents to
themselves and others! Who will help you if you become injured? If you work after regular
hours, find a "buddy" who will periodically check on you.
Only perform authorized procedures.
The storage and consumption of food, beverages, cosmetics, smokeless tobacco is limited to
"clean areas" outside of the laboratory.
Wash hands carefully before leaving the laboratory. Beware of contamination on clothing, door
knobs, frames, etc. Remove protective gear (gloves, etc.) before leaving the laboratory,
using a telephone, etc.
D. CHEMICAL SAFETY
1. GENERAL REQUIREMENTS FOR A SAFE LABORATORY
Chemicals should be handled in such a manner as to reduce the risk of personal exposure and
environmental damage. Some of the chemicals used in laboratories are acutely toxic
(causing asphyxiation, dizziness, neurotoxicity, allergic response, irritation, or body
corrosion), some cause chronic toxicity (mutagens, teratogens, carcinogens, compounds
causing reproductive toxicity, or other specific organ effects), and others may be flammable
or otherwise injurious. A list of the chemicals in the laboratory can be found in the Chemical
Inventory.
2. STANDARD OPERATING PROCEDURES FOR WORK WITH CHEMICALS:
a. Follow safe laboratory practices!
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Safe practices and procedures must be followed. Before starting any procedure, consider what
chemicals you are using and what safety procedures are necessary. Safety information is
available from several sources, including this document. For example, labels on chemical
bottles often contain a description of hazards associated with the chemical. Additional
information on safety procedures and signs/symptoms associated with chemicals can be
obtained by reading Material Data Safety Sheets, which are available at EH&S (tel. 2-5528).
The publication "Occupational exposure to hazardous chemicals in laboratories" is also
available at EH&S. The Dept. of Plant Pathology has also prepared a "Chemical Hygiene
Plan" which is available in the departmental office during business hours.
While the above sources will indicate the known hazards of chemicals, for caution's sake
consider any chemical a potential hazard!
b. Use protective and other safety equipment!
Safety Glasses should be worn to guard against flying particles, chemical splashes, etc. Larger
shields (such as splash goggles for face shields) should be utilized as necessary. Contact lens
should not be worn since they are extremely difficult to remove in case of chemical contact
with the eye, and may allow dangerous chemical vapors to collect behind the lens. If you
must wear contact lenses, inform your supervisor so that precautions can be taken.
Dust masks provide an extra although limited margin of protection when working with dry
chemicals. N95 style masks are available in the laboratory; complete Appendix D to Section
5144 of Cal/OSHA and return to Howard Judelson.
Chemical fume hoods provide a safe environment for working with chemicals yielding
hazardous vapors, dusts, and mists. Leave the sash (sliding door) at or below the safety
arrow to ensure adequate air flow and check the air flow before use.
Gloves protect the hands against chemicals, abrasion, heat, and cold. Before use, check gloves
for cracks, holes, etc. Do not touch door knobs, light switches, telephones, etc., with gloves.
Gloves vary in their resistance to different chemicals and physical conditions. Some
chemicals will dissolve some types of gloves! Gloves made of cotton or leather protect
against abrasion, sharp objects, glass, and in some cases, heat; however, they offer virtually
no wet chemical protection, and may actually absorb chemicals. Some rubber gloves may
contain pores that allow the passage of small chemicals, tritium, etc.
Lab coats and aprons protect the body. As with gloves, choose the material that best meets your
needs. Tyvek coats provide excellent general protection against chemicals and, unlike cloth
coats, do not absorb hazardous materials. Heavy duty rubber aprons provide good general
protection but are not resistant to all chemicals. As with gloves, lab coats and aprons should
remain in the laboratory. Many of the substances in the laboratory can be inadvertently taken
home on lab coats and aprons and are dangerous to friends, family, or pets.
Note: if you are sitting down, your lab coat may open up, exposing your legs or lap. As
appropriate, wear an apron that does not open up when you sit (for example, if you are sitting
by the chemical hood when dispensing a chemical).
Shoes with closed toes should be worn in the laboratory at all times to protect against spills and
splashes which reach the floor. Leather shoes offer better protection against corrosion than
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canvas shoes; open-toed shoes and bare feet are prohibited in the laboratory. If you must
wear sandals, then you must wear Tyvek shoe covers. Pets are not allowed in laboratories,
since they generally do not wear shoes and for other reasons, also.
Bench paper provides a way to protect your work area from contamination.
c. Handle and store chemicals safely!
In order to know how to safely handle chemicals, laboratory workers must be familiar with the
hazard characteristics associated with each chemical they use. This information is available
from several sources, including the bottle labels, Merck Index, EH&S, and Material Data
Safety Sheets (MSDS's, available at EH&S). MSDS's can be accessed through the EH&S
website (www.ehs.ucr.edu).
Make sure that others in the lab know the hazards associated with chemicals that you are using or
storing! Label your personal stock bottles or aliquots with "FLAMMABLE, CORROSIVE,
OXIDIZER, TOXIC, CARCINOGEN, etc." as appropriate, plus your name.
Never taste or smell a chemical, store food in a chemical container, or vice versa.
Store chemicals only on shelves with earthquake guards.
Often people think they are safe, if they are handling small amounts of a hazardous chemical.
However, you should realize that even if you are handling a few mls or grams of a
hazardous material, you may be taking it from a larger stock container that may pose a
major hazard if spilled.
d. Understand the different types of hazardous chemicals!
Organics, including solvents
The majority of chemicals in this class are toxic if inhaled or ingested, and many are
flammable. Some cause burns and others are carcinogens. Therefore, protective gloves and a
lab coat must be worn when handling them. As a general rule all organic solvents used in the
lab must be used only in the fume hood. The only exceptions are ethanol and isopropanol
which can be handled outside the hood (volumes less than 500 ml) if adequate ventilation is
available. Examples of hazardous organics are listed below.
Phenol is readily absorbed through the skin. Small amounts cause burns and large
amounts can cause death! Use a fumehood for phenol extractions. Make sure your wrists
as well as your hands and arms are covered. If the burn covers more than 10 cm2, or an
eye is affected, seek medical treatment.
PLEASE read the SOP document that describes detailed methods for dealing with phenol
spills on skin, including how to rinse skin with PEG or isopropanol solutions to extract the
phenol. THERE IS NO good method to rinse phenol from eyes; water is relatively
ineffective, but better than nothing. It is best to wear appropriate face or eye protection to
shield your eyes from a spill!
Diethyl ether and acetone are highly volatile, flammable, and explosive. Use in the
hood! Ensure that there are no open flames or sparking electrical devices present.
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Diethylpyrocarbonate (DEP) is used to kill enzymes. It can also kill you! Undiluted
DEP can cause severe skin burns, and the vapor can cause burns and irritation to the mouth
and nose. Always handle DEP in the fume hood. Always wear gloves and eye protection.
Exercise great care when carrying bottles of DEP about the laboratory.
Dimethylsulfoxide (DMSO) itself is not very hazardous, but it is frequently used as a
solvent for many dangerous chemicals. Since DMSO is almost instantly absorbed through
the skin, solutions employing DMSO must be handled with extreme care!
Strong Acids and Alkalis
Sulfuric acid, hydrochloric acid, chromic acid, glacial (100%) acetic acid, phosphoric acid,
and other strong acids can cause severe burns to the skin and their vapors can cause burns and
irritation to the mouth and nose. Handle concentrated acid solutions in the fume hood.
Always wear gloves and eye protection. Carry the stock bottles of these acids in the
protective carrier provided in the laboratory. Make acid solutions by adding the acid to water
(an alphabetic reminder: acid comes before water). Sulfuric acid and chromic acid are also
strong oxidizing agents and can cause explosions if mixed with organic solvents. Therefore,
store them away from organics. For the same reason, don't mix mineral acids with organic
acids (e.g. acetic acid). Solutions of sodium hydroxide, potassium hydroxide, and other bases
are also used in the lab, and these can cause severe burns. Always use gloves and eye
protection when handling these solutions. Store strong acids away from strong bases.
Oxidizers and Reducers
Examples of oxidizers include halogens, peroxides, potassium permanganate, ammonium
persulfate, and perchloric acid. Examples of reducers include ammonia, most organics,
carbon, and metals. Never store oxidizers with reducers, and keep oxidizers away from
flammables.
Carcinogens
Many carcinogens require special handling, and in some cases regulatory approval, before
their use. Bottles containing carcinogens (including personal stocks) must be specially
labeled. Some carcinogens deserve special mention:
Ethidium bromide is used in the laboratory as a stain for DNA and RNA. Always wear
gloves and a lab coat when handling this chemical. Do not dispose of ethidium bromide
solutions down the sink. To reduce spills and contamination, the stock solution is kept in a
special tray, along with a special pipettor. Only handle ethidium bromide solutions above
this container (to capture spills and drops).
Similarly, gel units (containing EtBr-containing gels) are operated in special trays so that
spills can be contained. Please wipe up any spills that occur outside the tray; spills in the
tray should be rinsed down the sink (see below).
Handing ethidium bromide waste: Very low concentration solutions, like used gel buffers,
can be disposed of down the regular sink in the electrophoresis area (do not use the hand
washing sink, please). For gel staining solutions, i.e. a low concentration solution, dispose
in the disposal bucket provided in the lab. High concentration solutions (>10 µg/ml), such
as CsCl gradients or spills of the stock solution, as well as contaminated solids, should be
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packaged separately and disposed of through EH&S. Gels containing ethidium bromide
are collected in a specially designated bucket.
Formaldehyde (formalin) is volatile and therefore represents a special hazard. It is also
flammable and irritates the eyes, nose, respiratory tract, and skin. All work with
formaldehyde must be done in a fume hood, or under the snorkel in the electrophoresis
area (for running formaldehyde-containing gels). Never pour formaldehyde down a sink.
Acrylamide and bisacrylamide are neurotoxins and carcinogens and can be particularly
dangerous when handled as a powder (usually we order it as a liquid, which is safer).
Gloves, lab coats, and masks must be worn when being weighed out. Warn people around
you that you are about to start weighing them out!
Chloroform is flammable and carcinogenic. Dispensing and handling should be
performed in a chemical fume hood.
Flammable Chemicals
Only store flammables in specially designed refrigerators or freezers; otherwise, sparking by
the motor or switches may cause an explosion. No more than 10 gallons of flammables may
be kept in each lab room. Do not shore flammables with oxidizers or acids.
Special instructions for the use of alcohol for flame-sterilization:
- alcohol for flaming should be kept in a flameproof container (no plastics!).
- only fill the alcohol reservoir with the minimum amount needed for sterilization
- always have a flameproof lid at hand to cover the reservoir in case it catches on fire. In case
of fire, DO NOT carry the flaming reservoir to the sink or any other location! Cover it with
the lid and let it burn out.
- make sure that the reservoir lid is kept in a location that will not be occluded by flames.
- plan your movements to ensure that the reservoir of alcohol is far enough from the flame
such that flaming alcohol can not fall into the reservoir.
- if you catch on fire, fall to the ground and roll. Also use the fire blanket to extinguish the
flames.
Toxic chemicals.
Examples include cycloheximide, hygromycin, benomyl, and acrylamide. When using dry
forms of toxic chemicals, work in a fume hood, wear a protective mask, gloves, and a lab coat.
Warn those around you when you are about to use them. If you ingest any of these chemicals,
seek medical attention - symptoms may not appear immediately.
Radioactive compounds
A variety of radioactive compounds are used in the laboratory, which can pose a hazard if
improperly used. See section E for more information.
Compressed Gases
Compressed gases used in laboratories include nitrogen, argon, and others. Their primary
dangers are asphyxiation and explosions. Do not use gas cylinders unless you have been
trained in their use. Make sure cylinders are firmly anchored to a wall, cabinet, or cylinder
cart, and protect the cylinders from dropping, knocking, or otherwise shocking. Always use in
a well ventilated area.
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Cryogenic materials
Liquid nitrogen and dry ice can cause severe cold burns and asphyxiation. Take the following
precautions:
(i) Do not withdraw liquid nitrogen from stock tanks unless you have been shown how to do
so safely.
(ii) Use a suitable container to carry liquid nitrogen. Plastic or stainless steel dewar flasks are
suitable. Styrofoam ice buckets, glass thermos flasks, plastic or glass beakers are not safe
containers!
(iii) Wear protective clothing when working with liquid nitrogen - i.e. lab coat, protective face
mask or goggles, suitable insulated gloves (autoclave gloves are OK), adequate footwearno sandals etc. Wear gloves when using dry ice.
(iv) Work in an adequately ventilated area.
(v) When done, dump out unused liquid nitrogen outside. Unattended cyrogenics can spill on
the unsuspecting and cause injury.
e. Know what to do in case of a chemical spill!
Alert personnel in the immediate vicinity.
If the spill poses a threat to you or others (i.e. a volatile hazardous compound), hold your breath,
evacuate the area, and close all doors. Pull a fire alarm to evacuate the building. Once you
get to a safe area, call 911 to report the nature of the incident.
If the spill is small (500 mls or less), the laboratory SPILL CONTROL KIT can be used. Call
EH&S (x5528 or 5222) or 911 for larger spills. Put on protective equipment (gloves, eye
protection, lab coat). Absorb spilled liquid in vermiculite. Neutralize acids and bases
using sodium bicarbonate. Scoop absorbent into plastic bags and dispose of properly.
If someone has been splashed by a wet chemical, flush the affected area with water, using safety
shower or eyewash when appropriate. In the case of dry chemical contact, first brush off
the chemical before washing with water. In case of eye contact, hold the eyelids open,
wash the eyes for 15 minutes while rolling the eyes to achieve good flushing, and then get
medical assistance.
f. Chemical waste must be properly disposed of!
EMPTY CHEMICAL CONTAINERS must be rinsed three times with water before disposal (in
the regular trash).
LIQUID AND SOLID WASTE must be disposed of in leak-proof containers. The containers
should be labeled with the date the first items were added (using a label from the on-line
waste tag program), and with the chemical nature of the contents. Liquid waste bottles
must have screw-caps and head-space should be provided for expansion. Do not stockpile
waste in the lab, and do not let wastes age and become reactive (for safety dispose of
wastes more than 6 months old. Let Howard Judelson know when waste containers need
to be picked up by EH&S.
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SOLIDS CONTAMINATED WITH CHEMICALS must not have liquid in them. They must be
placed in a sturdy plastic (i.e. leakproof) container, and disposed of through EH&S.
Plasticware contaminated with small amounts of volatile chemicals (i.e. chloroform) can
be kept in the hood until the liquid evaporates, and then disposed of in the regular trash.
DO NOT LET THE WASTE CONTAINER OVERFLOW. Let Howard Judelson know when a
container is about 80% full.
E. RADIOACTIVE COMPOUNDS
DO NOT work with radioactivity until you have obtained specific authorization from Howard
Judelson and have been placed on the laboratory license. Follow these guidelines when
working with radioactivity:
1. Handle radioactivity only in authorized areas of the laboratory. A specific radioactive work
area, containing equipment dedicated to radioactive experiments, is provided so DON'T
WORK WITH RADIOACTIVITY ANYWHERE ELSE!
2. Wear the proper personal protective equipment: gloves (double-layered) and lab coat. Don't
wear open-toed shoes or sandals. Cover up your legs with a long coat if you are wearing
shorts. Always wear eye protection, especially when working with high-energy particles
such as 32P (eyes are very sensitive to irradiation). When working with tritiated
compounds, change your gloves frequently since tritium seeps through latex and vinyl
gloves!!
3. Before starting your work, sign up on the area use log to indicate the isotope, procedure,
and area that you are working in.
4. Always handle radioisotopes on absorbent bench paper.
5. Always minimize your exposure to radioisotopes! Use the proper shielding to protect you
and anyone else in the room. Minimize the amount of time that you hold radioactive items
or stand in the work area. Try to use less isotope when possible.
6. All unattended radioactive items (experiments in progress, materials stored in the freezer,
refrigerator, etc.) MUST be labeled with the isotope, date, and your initials. This includes
scintillation vials!!
7. Dispose of solid and liquid waste in the proper containers. NO radioactivity should go in the
normal trash or down the sink.
8. Prevent the proliferation of radioactivity!! Once you have handled anything in the radioactive
area remove your gloves before touching anything outside the area. Only use the
radioactivity-dedicated Pipetman for work with isotopes. Avoid bringing in more racks,
ice-buckets, etc. into the radioactivity area: once it is there it must be assumed radioactive!
9. CLEAN UP ALL SPILLS! Cleaning solutions are provided in the radioactivity room. For
contaminated skin: use only the denoted hand cleaner (others are caustic and will eat away
your skin, speeding the transfer of the isotope into your bloodstream) and don't scrub hard
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(your outer layer of skin provides a barrier against the ingress of chemicals into your
body).
10. FILL OUT THE LOG SHEET! All shipments of radioactivity received in the lab, and all
withdrawals of radioactivity from the stock bottles, must also be recorded on the log sheet.
11. When done with your work, check your area for contamination. Also, clean up any debris
(paper towels, graduated cylinders, pens, etc.): store them in the proper places or discard
as appropriate. YOU ARE RESPONSIBLE FOR LEAVING YOUR WORK AREA
SPOTLESS!!!! If the area was messy before you started, clean it up by either doing it
yourself or contacting the previous person to work there.
F. PHYSICAL HAZARDS
1. GLASSWARE
Glassware can break unexpectingly. Inspect for cracks or sharp edges before use. Dispose of
all glass in the special glass waste containers. Throw out broken glassware: it is cheaper to
buy a new beaker than a new finger. Hand protection should be used when picking up
broken glass.
Be cautious when stressing glass. This includes moving glass between temperature extremes,
inserting tubing into stoppers, creating a vacuum within glassware, etc.
2. NEEDLES, SYRINGES, RAZOR BLADES
Handle these items with special caution. Leaving these items in common areas is reckless and
inconsiderate. Dispose of these items in the special "sharps" container present in each lab.
3. HEATING BLOCKS AND BURNERS
Never leave these items unattended! Label hot plates as "HOT" when you are finished using
them and waiting for them to cool. Operate flames in a safe and uncluttered environment. It
is dangerous to operate flames above paper lab mats and near flammable liquids; exercise
extreme caution when flaming tools with ethanol.
4. ULTRAVIOLET (UV) LIGHTS.
Ultraviolet lights are used in biological safety cabinets to sterilize the interior, and to illuminate
nucleic acids stained with fluorescent dyes. Your eyes are most sensitive and you can
receive permanent damage to your retinas. The skin is also sensitive depending on whether
you have a fair or dark complexion, and you can receive burns (literally sunburn) from
extended exposure. Both short and long wavelengths cause damage.
When using a biological safety or sterile hood, ensure that any UV lights are off. Never look
into a cabinet when the UV light is on.
When illuminating DNA with UV light, wear UV protective goggles. Wear a face mask if you
are going to be facing directly into the light for more than a few seconds. If you are going to
be exposed to the UV light for more than 5 minutes, you should cover exposed skin with
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clothing or a blockout-type sunscreen. If you have a very fair, easily-sunburnt complexion,
you should observe these precautions for any exposures.
5. ELECTRICAL EQUIPMENT
Electrical circuits can kill. Tests at the Howard Hughes Institute indicate that even common
bench-top gel boxes running at 100 volts can be lethal under some conditions.
Do not use an electrophoresis apparatus unless you have instructed how to do so safely. Do not
defeat or remove safety interlocks. Never touch, interfere with or move an operating
electrophoresis apparatus. This particularly applies to the high voltage apparatuses used for
DNA sequencing. Always ensure that there is an adequate amount of buffer in the
electrophoresis tanks to prevent arcing. Never leave a gel unattended if there is any question
that the tanks may be leaking. Do not apply excessive voltages to agarose gels as the gels
may melt, dry out, and eventually catch fire.
Only experienced researchers who have been trained to work safely with line voltage circuits
(i.e. 120, 230 volts) may disassemble such electrical equipment.
6. CENTRIFUGES.
A variety of centrifuges are encountered in the lab, from small bench centrifuges to
ultracentrifuges. The primary risks from centrifuges are physical injury due to the high speed
and forces generated. Back injuries can also be caused by trying to lift heavy rotors. Observe
the following precautions:
(a) Do not use any centrifuge or rotor unless you have received instruction on how to use
it. This especially applies to the ultracentrifuges.
(b) If a centrifuge has a door, keep it closed during operation - NEVER bypass a door
safety lock in order to open an operating centrifuge.
(c) If a centrifuge has no cover, keep well clear during operation.
(d) NEVER USE YOUR HANDS TO SLOW A CENTRIFUGE ROTOR!
(e) When lifting a heavy rotor, especially for the first time, test its weight cautiously before
attempting to move it. Do not attempt to lift it unless you are able to lift twice the
weight. If unsure seek assistance.
(f) Never move a spinning centrifuge!
7. OVENS, AUTOCLAVES AND MICROWAVES.
The key danger here is burns and scalds. Damaged microwave ovens can also emit dangerous
microwaves. Observe the following precautions:
(a) Always wear protective gloves when removing items from an autoclave or oven.
(b) If the items are large, heavy or bulky, seek assistance (and/or use a cart).
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(c) If you leave hot items on a bench unattended, leave a note alerting others to the fact that
they are a burn hazard.
(d) When removing hot liquids from an autoclave or microwave oven, beware that the
liquid may be superheated and may boil over vigorously if agitated. This is a
particular problem with viscous liquids such as agar and agarose solutions. If in doubt,
do not lift the container until you are sure it will not boil over when you pick it up.
Instead agitate it several times without picking it up - very gently at first then with
increasing vigor.
(e) Never lean into an autoclave immediately after you have opened it - the escaping steam
can cause you a severe facial burn. Wait a few moments until the steam has dissipated.
G. BIOLOGICAL AGENTS: PROCEDURES AND PRECAUTIONS
1. GENERAL CONSIDERATIONS
Various organisms are used in the laboratory, including fungi (Phytophthora infestans and
Saccharomyces cerevisiae), bacteria (Escherichia coli), and plants (tomato, potato,
petunia).
There is no danger of you becoming infected with these organisms as they do not cause human
diseases. For organisms containing recombinant DNA, the main precaution is to prevent
their escaping into the general environment. Many of the isolates of Phytophthora used in
the laboratory are not native to California and have been imported under special licenses
from the United States Department of Agriculture. These must also be contained within
the laboratory environment.
2. OPERATING PROCEDURES
a. General work procedures
Standard good microbiological practices, plus special procedures to maintain containment
of exotic organisms and organisms containing recombinant DNA, are absolutely required.
(a) Access to the laboratory will be limited at the discretion of Howard Judelson.
(b) Work surfaces must be decontaminated at least once a day with 70% ethanol. The sites
of minor spills (<5 ml) should be decontaminated with 0.5% (w/v) sodium
hypochlorite.
(c) Pipetting must be carried out using only mechanical aids. No mouth pipetting!
(d) Eating, drinking, smoking and applying makeup are not allowed in the work areas of
the laboratory. Food may be stored only in refrigerators reserved for that purpose
located in the offices or the lunch room.
(e) The use of a laboratory coat is a good practice when handling microorganisms. Keep
these coats in the lab; do not wear them outside!
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(f) The recirculating sterile hood (a class 2 biological safety cabinet) must be used as a
protective measure against spread of the organisms within the laboratory when handling
cultures that are producing airborne spores (e.g. Phytophthora infestans).
(g) Windows and doors to the outside should not be left open. The lab should be locked
when unattended.
(h) Wash your hands before leaving the laboratory, if you have been handling viable
cultures (or hazardous chemical reagents).
(i) Freezers, refrigerators and incubators used for storage of recombinant organisms must
be labeled with the biohazard symbol.
b. Disposal of biological waste and decontamination of equipment
Liquid waste. All liquid cultures, culture fluids, spore or cell suspensions, pot runoff water from
infected plants etc. must be autoclaved for at least 20 minutes (40 minutes for volumes
greater than 2 liters) before being poured down the sink. Alternatively, bleach can be
added to 10%, and incubated for at least one hour.
Please think of your labmates when using bleach. A container with bleach should be
covered and labelled.
Solid waste. All solid material including agar plates, paper, gauze, etc. must be sealed in two
strong plastic autoclave bags, and then autoclaved for 40 minutes with indicator tape.
Materials contaminated with transgenic plant pathogens or with airborne propagules must
be sealed within a watertight lightweight plastic bag before being placed in the biohazard
bags to ensure that spores, contaminated moisture etc. cannot escape prior to sealing of the
autoclave bags. Contaminated sharp items, e.g. razor blades, scalpel blades, needles,
disposable glassware etc. must be placed in a puncture-resistant, leak-proof container.
When full the boxes must be autoclaved for 40 minutes.
Contaminated Hardware and Glassware. These must be autoclaved for 20 minutes or soaked in
10% bleach for 30 minutes prior to washing. Glass pipettes must be placed in a solution of
5% bleach for at least sixteen hours at room temperature prior to washing. If the items are
contaminated on the outside they must be placed in an autoclavable tub prior to
autoclaving. Non-autoclavable items, e.g. some plasticware, polycarbonate centrifuge
tubes, homogenizers etc. must be soaked in 10% bleach prior to washing.
c. Spills and emergency procedures
Spills: The prime priority is to contain the organisms. Liquid spills should be wiped up with
absorbent paper or cloth and the contaminated material placed in an autoclave bag for
sterilization. Wipe area with 5% bleach. Solid materials should be carefully enfolded in
absorbent paper or cloth and then autoclaved. Cultures with airborne propagules should be
contained by gently laying cloth or paper soaked in 5% bleach before further handling.
Please notify Howard Judelson of any spills of greater than 10 ml (home phone 686-9323).
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Ingestion, inhalation, self-inoculation, etc. None of the organisms are infectious to humans. In
the case of self-inoculation (e.g. with an infected needle) or large ingestion of E. coli
strains containing vector plasmids or recombinant DNA, treatments involving antibiotics to
which the plasmids confer resistance (e.g. ampicillin, tetracycline, chloramphenicol,
kanamycin) should be avoided for one week to minimize the chance of the gut being
colonized or the plasmids being transmitted to enteric bacteria.
Natural disasters, fires, power failure. None of the organisms is infectious to humans so there is
no danger to emergency personnel. Release to the environment is to be avoided because
they may harbor recombinant DNA, but there would be no direct or immediate hazard to
the local population or to the local flora and fauna. If there is a disaster, fire or power loss
while cultures are being handled in a class ll Biological Safety Cabinet, close the front
shield on the cabinet. In the case of rupture of culture vessels following a natural disaster
(i.e. an earthquake) or fire, the spilled cultures should be cleaned up as soon as practical,
following the instructions listed above for spills.
I. SAFE USE OF COMPUTERS
When working on the computer for long hours, pay attention to tension, discomfort or pain you
feel and take immediate action to relieve it.
Make sure you stand up and walk away from your computer on a regular basis. This should be
done at least every hour, for a few minutes. Also remember the "rule of 20" to rest your
eyes: every 20 minutes, focus your eyes on something 20 feet away for 20 seconds.
The greatest risk for problems occurs when people use computers intensely for long hours and
work in poor postures. To minimize problems, adjust you chair so your feet and back are
firmly supported by the floor and seat back, and centered in front of the computer
J. OTHER SAFETY-RELATED DOCUMENTS
In addition to the resources available through EH&S, you should be aware of the location of
other documents stored in the Judelson lab. These include (i) the departmental Chemical
Hygiene Plan, and (ii) Standard Operating Procedures, i.e. SOPs for hazardous operations.
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I. ATTESTATION OF TRAINING
This document has provided information on procedures and policies required for safe laboratory
practices, and information on sources of additional information regarding workplace
safety. Please keep it for your reference.
By your signature, you are acknowledging that you have received safety information and
instruction regarding the above items. Specifically, you are certifying that you have been
informed, in general, of (a) potential occupational hazards in the work area and associated
with your job assignment; (b) safe work conditions, practices, and protective equipment
required for job activities; (c) hazards of chemicals to which you may be exposed and your
right to information contained in material data safety sheets for those chemicals; (d) your
right to ask questions or to provide any information to the employer on safety matters
either directly or anonymously without any fear of reprisal; and (e) that compliance with
safe work practices are required for all students or academic or staff researchers. Please
return this page to Howard Judelson.
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Employee name (printed)
(Employee signature)
Date