Download Dr. Tim Chataway

Safe Work Method Statement
Acrylamide Gel Electrophoresis
SWMS NUMBER
RISK ASSESSMENT
NUMBER
RISK ASSESSMENT
Medium
DATE CREATED /
REVIEWED
15.07.13
Employees Involved in SOP
Creation
Dr. Tim Chataway, Angela Binns
CONTACT PERSON
Dr. Tim Chataway
Personal Protective Equipment Required
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Laboratory coat.
Covered shoes
Mask to be used when weighing the chemical SDS.
Gloves – to be worn throughout entire procedure.
Eye protection as required
Before work Commences
 Users must be trained and training documented.
 Refer to the SDS for each chemical
 Only use commercially made Acrylamide (DO NOT make your own use only pre-made)
 Read and understand the procedure.
Hazards
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Hazardous substances
Electric shock – electrophoresis power supply.
Lacerations
Needle-stick injury
Eye damage
1. Reagents
Acrylamide solution
Tris buffers
SDS solution
Ammonium Persulphate
TEMED
Sample buffer
The hazards are listed on the SDS for each chemical and are listed on Chemwatch. Essentially,
gloves must be worn throughout this procedure. If the gloves are contaminated with one of the
chemicals, the gloves must be changed or rinsed thoroughly in water before proceeding. A mask
must be worn when weighing the chemical SDS.
Solutions
Acrylamide
Acrylamide solutions should not be made from dry powder. Acylamide is a cumulative neurotoxin
and the dry powder is hazardous. There are commercially available pre-made solutions which
should be used.
Tris buffers
Tris buffers do not pose any significant hazard. However, they need to have their pH adjusted
using strong acids or bases. Gloves and eye protection must be worn. Read the SOP for using a
pH meter.
Equipment
Electrophoresis involves the use of power supplies which produce lethal levels of electricity.
However, the risk is negligible if safe procedures are followed. Ensure that the leads are inserted
correctly, all fluids are kept away from the power supply and the electrophoresis tank is assembled
with its lid in place before attaching the power supply.
Procedure
Clean glassware with Extran
solution
Remove adherent acrylamide
with razor blade
Assemble plates and spacers
into gel pouring apparatus
Make up acrylamide solution
of required percentage
Degas solution
Add catalyst, pour solution
into plate apparatus, allow to
set
disassemble plates, insert
plates into electrophoresis
tank, add upper and lower
tank buffers
Prepare samples in SDS
loading buffer, heat at 95 deg
C in heating block
Load samples into gels
Apply voltage (200Volts)
Remove gels from tank,
separate glass plates, place gel
in tray for staining, add
Coomassie Blue
Destain with methanol/acetic
acid solution
Risk
This procedure involves potential
exposure to toxic chemicals
Eye injury or laceration if plates are
dropped
Finger laceration
Eye injury or laceration if plates are
dropped
Exposure to toxic chemicals
Precautions
Gloves must be worn for all
steps in this procedure
Eye protection, wear gloves,
covered shoes
Use single sided blade only,
apply at an oblique angle
Eye protection, wear gloves,
covered shoes
gloves
Explosive release of glass/acrylamide
Use glassware designed for
vacuum use
Burnt fingers, lid pops off if heat too
high
Do not touch block, ensure
correct temperature, label
heating block as being hot to
prevent others from
accidental burns
Load samples with 10 uL
pipette
Ensure leads connected
correctly, correct buffer and
fluid levels, lid in place on
tank before applying voltage
Eye protection, gloves
Needlestick injury if using needle and
syringe
Electric shock
Injury from glass
Eye injury from acetic acid
Use with care, eye
protection