Cleaver Scientific omniPAGE Maxi User manual

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omniPAGE Maxi and
Mini Wide Vertical
Electrophoresis Units
Instruction Manual
Catalogue Numbers
VS20WAVESYS
VS20WAVESYS-CU
VS20WAVED
WAVETETRAD
VS10WD
VS10WDSYS
VS10WDSYS-CU
Record the following for your records:
Model _____________________
Catalogue No. _____________________
Date of Delivery _____________________
Warranty Period _____________________
Serial No. _____________________
Invoice No. _____________________
Purchase Order No. _____________________

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Contents
Instruction Manual 1
Catalogue Numbers 1
Contents 2
Safety Information 3
Packing List 4
Specifications 6
Operating Instructions 7
Usage Guidance and restrictions 7
Setting up the omniPAGE Gel Tanks 7
Fitting Electrode cables 7
Casting Unit Preparation 7
Cleaning the Glass Plates 7
Gel Cassette Assembly 8
Casting Stand Assembly 8
Casting 4 gels 9
Gel Preparation 12
Gel Volumes 12
Gel Mixture 13
Gel Pouring 13
Sample preparation and Loading 14
Denatured sample preparation 14
Loading the samples 15
Buffer Volumes 16
Gel Running 16
Connecting a recirculating chiller 17
Solutions (For SDS-PAGE) 17
References 18
Troubleshooting 19
Care and Maintenance 22
Cleaning Vertical Units 22
RNAse Decontamination 22
Ordering information 23
Comb options 24
Related Products 25
Warranty 27

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Safety Information
When used correctly, these units pose no health risk. However, these units
can deliver dangerous levels of electricity and are to be operated only by
qualified personnel following the guidelines laid out in this instruction manual.
Anyone intending to use this equipment should read the complete manual
thoroughly. The unit must never be used without the safety lid correctly in po-
sition. The unit should not be used if there is any sign of damage to the exter-
nal tank or lid.
These units comply with the following European directives:
2014/35/EU Low Voltage Directive and 2014/30/UE (official Title 2004/108/EC)
EMC Electromagnetic Compatibility
By virtue of the following harmonised standards:
BS EN IEC 61010-1: 2010 Safety Testing of Lab Equipment
BS EN IEC 61326-1:2013 EMC Electro Magnetic Compatibility
ROHS DIRECTIVE 2011/65/EU
BS EN 50581:2012 Restriction of Hazardous Substances

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Packing List
Each omniPAGE WAVE Maxi unit includes a tank, wired PAGE module, lid,
CSL-CAB cables, VS20WAVE-CC Cooling coil and the following items:
SKU
Glass Plates
Combs
Caster
VS20WAVESYS
VS20NG –Notched, Pk/2
VS20PGS1 –Plain with bonded
1mm spacers, Pk/2
VS20-DP –Dummy Plate
2 x VS20-24-1
1mm thick, 24
sample
VS20DCAST -Caster
VS20WAVESYS-CU
As in VS20WAVESYS
As in VS20WAVESYS
VS20DCAST -Caster
VS20WAVE-EC –addi-
tional casting stand
without electrodes
VS20WAVED
As in VS20WAVESYS
As in VS20WAVESYS
None
WAVETETRAD
1 of VS20NG –Notched, Pk/2
2 of VS20PGS1 –Plain with
bonded 1mm spacers, Pk/2
1 of VS20NGS1 Notched with
1mm spacer, Pk/2
VS20-DP –Dummy Plate
4 of VS20-24-1
1mm thick, 24 sam-
ple
VS20DCAST -Caster
VS20WAVE-EC –addi-
tional casting stand
without electrodes
Each omniPAGE Mini wide unit includes a tank, wired PAGE module, lid, CSL-
CAB cables, VS20ICB Cooling pack and the following items:
SKU
Glass Plates
Combs
Caster
VS10WD
VS10WNG - Notched, Pk/2
VS10WPGS1 –Plain with bonded
1mm spacers, Pk/2
VS10W-DP –Dummy Plate
2 of VS20-24-1
1mm thick, 24 sam-
ple
None
VS10WDSYS
VS10WNG - Notched, Pk/2
VS10WPGS1 –Plain with bonded
1mm spacers, Pk/2
VS10W-DP –Dummy Plate
2 of VS20-24-1
1mm thick, 24 sam-
ple
VS20DCAST
VS10WDSYS-CU
VS10WNG - Notched, Pk/2
VS10WPGS1 –Plain with bonded
1mm spacers, Pk/2
VS10W-DP –Dummy Plate
2 of VS20-24-1
1mm thick, 24 sam-
ple
VS20DCAST
VS10WEXCASTER –addi-
tional casting stand with-
out electrodes
Packing List Checked by: ________________________
Date: ________________________

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The packing lists should be referred to as soon as the units are received to
ensure that all components have been included. The unit should be checked
for damage when received.
Cleaver Scientific is liable for all missing or damaged parts / accessories within
7 days after customers have received this instrument package. Please contact
Cleaver Scientific immediately regarding this issue. If no response within such
period is received from the customer, Cleaver Scientific will no longer be liable
for replacement/damaged parts.
Please contact your supplier if there are any problems or missing items.

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Specifications
1
Lid and Cables
1500V Rated cables, high impact acrylic lid
2
PAGE Insert
Glass filled Nylon and Acetal construction
3
Clamp Screws
Injection moulded to avoid thread wear
4
Plate Clamps
Injection moulded, provide even pressure across glass
plates
5
Glass plates
4mm thick float glass
6
Inner Buffer Chamber
Allows use of separate inner running buffer
7
Electrodes
99.99% pure platinum electrodes
8
Cooling Coil
Can be connected to recirculating chiller
9
Out Buffer Chamber
Injection Moulded High Impact Acrylic
10
Casting Base
Injection Moulded Acetal
11
Casting Mat
Ultra-soft silicone for leak free casting
12
Combs
Lapped Acrylic for high thickness uniformity, also availa-
ble in polycarboante

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Operating Instructions
Further information (including videos) regarding setting up and running the
omniPAGE units can be found at www.cleaverscientific.com
Usage Guidance and restrictions
•Maximum altitude 2,000m.
•Temperature range between 4°C and 65°C.
•Maximum relative humidity 80% for temperatures up to 31OC decreas-
ing linearly to 50% relative humidity at 40OC.
•Not for outdoor Use.
This apparatus is rated POLLUTION DEGREE 2 in accordance with IEC 664.
POLLUTION DEGREE 2, states that: “Normally only non-conductive pollution
occurs.
Occasionally, however, a temporary conductivity caused by condensation
must be expected”.
Setting up the omniPAGE Gel Tanks
Fitting Electrode cables
•Note the position of the lid on the unit. This shows the correct polarity
and the correct orientation of the cables, black is negative and red
positive.
•Remove the lid from the unit. Note if the lid is not removed, fitting the
cables may result in un-tightening of the gold plug and damage to the
electrode.
•Screw the cables into the tapped holes as fully as possible so that there
is no gap between the lid and the leading edge of the cable fitting.
•Refit the lid.
The unit is now ready to be used.
Casting Unit Preparation
Cleaning the Glass Plates
Clean a set of glass plates for each gel first with distilled water and then with
70 % ethanol. One set of glass plates constitutes one notched glass plate
and one plain glass plate with bonded spacers.

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When using a triple glass plate sandwich, two notched glass plates are re-
quired, one set of free spacers and a set of plain glass plates with bonded
spacers. The plain glass plate is positioned outermost, then a notched glass
plate, free spacers and second notched glass plate. Alternatively, accessory
notch glass plates with bonded spacers are available.
Note: All glass plates, gel casting modules, casting base and accessories must be com-
pletely dry before the set –up. Wet components are more likely to miss-align and cause
leaks.
Gel Cassette Assembly
Assemble the glass plates so that the bottom of the glass plates and the
spacers are perfectly aligned.
A triple plate sandwich can be used by combining the standard Notched
plate and spacer plate, with an additional notched plate with spacers. For
triple plate sandwiches, the free spacers Need to be perfectly aligned which
is best performed using a small spacer or comb to push the spacers apart.
Notched glass plates with bonded spacers do not need manual alignment.
NOTE: The glass plates with bonded spacers have an arrow in the top of the spacers which
are slightly longer than the glass plate to indicate the top.
Casting Stand Assembly
1. Ensure that the clamps are adequately open for
the thickness of spacer used. The clamps can be
opened by loosening the screws and sliding the
clamps.
2. Position the Slab Gel Insert on a flat surface. Insert
the glass plates into the Slab Gel Insert between
the clamps and the white gasket. The Slab Gel In-
sert contains clamps which impart even pressure
onto the glass plates and allow even screw pres-
sure transfer onto the sealing edge of the glass
plate, ensuring complete sealing.
3. Fully tighten the screws, making sure not to wob-
ble the unit.

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4. When only one gel is being run, the dummy plate
must be used in the second position and fully
tightened.
5. Position the Slab Gel Insert in the casting base
such that the Cam pins have handles pointing
downwards and are located in the insert holes.
The top of the module may need to be pushed
down very slightly to locate the cam pins.
NOTE: At this stage, check that the bottom edges of the spacers
and glass plates are perfectly aligned.
6. With the cam pin handles facing directly down-
wards, turn the cam pins fully through 135° or until
the insert has tightened onto the silicone mat.
NOTE: It is best to turn the cams in opposite directions to each
other. Do not overturn as this will cause the glass plates to push
upwards and the assembly will be more likely to leak. The unit is
now ready for gel preparation and pouring.
7. Pour in the gel solution, insert the combs and al-
low the wells to polymerise. Gel pouring is best
performed using a 25ml or 50ml pipette. If using a
stacking gel, don’t insert combs, allow gel to pol-
ymerise, pour stacking gel and then insert combs.
Casting 4 gels
The thicker green sliding gel clamps are recommended to secure up to 2 gels
(i.e. 1 gel either side of the PAGE insert) for gels up to a maximum thickness of
2mm. For 4 gels (i.e. 2 gels either side of the PAGE insert: made using 1 plain
glass plate and 1 notched glass plate, both with bonded spacers, and 1
notched plate without spacers), the thinner YELLOW sliding gel clamps must
be used.
To convert the unit from a 2- to 4-gel configuration please adhere to the fol-
lowing instructions. One completed, proceed with casting as above.

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1. To replace the green sliding gel clamps, begin by
unscrewing the colour-coded vertical screw pins.
There should be no glass plates within the PAGE in-
sert, in the side(s) being unscrewed. Once the screw
pins are unscrewed sufficiently the green clamp
should sit in the resting slot as shown.
2. Gently push the green sliding clamp horizontally to-
wards the core of PAGE insert until it can move no
further.
3. Push the green sliding gel clamp out of the PAGE
insert by gently pressing the protruding pin as shown.
4. Once the green sliding gel clamp is removed the
PAGE insert is ready to accept the thinner yellow
sliding gel clamps for 2-gel sandwiches, either side
of the PAGE insert, to convert the WAVE to a 4-gel
configuration.

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5. Insert the yellow sliding gel clamp into the hole clos-
est to the core of the PAGE insert. Once inserted
gently withdraw the sliding clamp outwards away
from the core of the PAGE insert as shown.
6. Withdraw the sliding gel clamps so that they sit sus-
pended in the resting slots as shown. Repeat steps
1-6 to replace the remaining green sliding gel
clamps. Once complete the PAGE insert is ready for
4-gel assembly.

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Gel Preparation
Care should be taken when selecting the pore size of the gel to be used. The
pore size or % of gel determines the resolving ability given different sizes of
protein.
Gel percentage for various protein sizes are shown below:
Acrylamide Percentage
Separating Resolution
5 %
60 - 220 KD
7.5 %
30 - 120 KD
10 %
20 - 75 KD
12%
17 –65 KD
15 %
15 -45 KD
17.5%
12 –30 KD
Gel Volumes
For omniPAGE Maxi
Number of Gels
Volume for 1mm Thick Gel
Single
35 ml
Double
70 ml
4 Gels (2 x Triple plate)
140 ml
For omniPAGE Mini Wide
Number of Gels
Gel Thickness (mm)
Volume (mm)
Single
0.5
8.75
1
17.5
1.5
26.3
2
35
Double
0.5
17.5
1
35
1.5
52.5
2
70
4 Gels (2 x Triple plate)
0.5
35
1
70
1.5
105
2
140

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Gel Mixture
Prepare gel solutions as per tables below. These give the volumes of solutions
from the standard stock solutions. These should be gently mixed avoiding
generation of bubbles which will inhibit polymerization by removing free radi-
cals. The following values are for a 1mm gel for the omniPAGE Maxi, adjust
volumes accordingly for other gel sizes.
Resolving Gel:
Solution
5 %
7.5%
10 %
12%
15 %
17.5%
Distilled Water
41ml
35.25m
29.6ml
24.7ml
17.6ml
11.7ml
30 % Stock Acrylamide Solu-
tion
11.7ml
17.6ml
23.5ml
28.2ml
35.25ml
41.1ml
4 X Resolving Tris Solution
17.6ml
17.6ml
17.6ml
17.6ml
17.6ml
17.6ml
Before pouring, add and mix 700 µl of 10% ammonium persulfate and 70 µl of TEMED, these are
the polymerising agents and will initiate polymerisation of the gel, so must be added only immedi-
ately before pouring.
Stacking Gel
Solution
VS20WAVE
VS10W
Distilled Water
16.8ml
8.4ml
30 % Stock Acrylamide Solution
2.6ml
1.3ml
4 X Stacking Gel Tris Solution
6.4ml
3.2ml
Before pouring, add and mix 178 µl of 10% ammonium persulfate and 17.8 µl of TEMED for an om-
niPAGE Maxi gel or 134 µl of ammonium persulfate and 13.4 µl of TEMED for an omniPAGE Mini
Wide gel. These are the polymerising agents and will initiate polymerisation of the gel, so must be
added only immediately before pouring.
Gel Pouring
With Stacking Layer
1. Insert the comb into the glass plates and mark a point on the glass
plates 1cm below where the comb teeth finish. This indicates where to
add the resolving gel to.
2. Add polymerising agents to the gel solution and mix well but avoid gen-
erating air bubbles.

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3. Fill the glass plates again avoiding generating any air bubbles. Filling
must be performed quickly before the gel becomes too viscous.
4. Overlay the gel carefully with 1 ml of Isobutanol, Isopropanol or distilled
water. When using distilled water extra care must be taken to ensure
there is no mixing with the gel solution.
5. Let the resolving gel polymerize. Usually this takes around 15 minutes,
but this can vary due to the freshness of the reagents used. If polymeri-
zation takes a lot longer than this, use fresher stock solutions or add
more APS and TEMED.
6. Carefully mix the stacking gel solution, avoiding generating air bubbles.
7. Pour off the overlay liquid from the resolving gel and rinse the gel with
distilled water.
8. Add polymerising agents to the stacking gel and pour over the resolv-
ing gel.
9. Carefully insert the comb making sure that no air bubbles get trapped
under the ends of the comb teeth as these will inhibit sample progres-
sion.
10. Allow the stacking gel to polymerize for 30 minutes.
With Stacking Layer
1. Add polymerising agents to the gel solution and mix well but avoid gen-
erating air bubbles.
2. Fill the glass plates again avoiding generating any air bubbles. Filling
must be performed quickly before the gel becomes too viscous.
3. Carefully insert the comb making sure that no air bubbles get trapped
under the ends of the comb teeth as these will inhibit sample progres-
sion.
4. Let the resolving gel polymerize. Usually this takes around 15 minutes,
but this can vary due to the freshness of the reagents used. If polymeri-
zation takes a lot longer than this, use fresher stock solutions or add
more APS and TEMED.
Sample preparation and Loading
Denatured sample preparation
The instructions given below are for denatured samples. For Native samples,
please consult a laboratory handbook.

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1. Prepare the protein samples for loading. The volume of sample de-
pends on the capacity of the wells (See Comb specifications page 28).
2. Using a 0.5 ml micro-centrifuge tube or other convenient receptacle,
combine the protein sample and 4 X sample buffer. It is always advisa-
ble to use protein markers in one of the end lanes to indicate sizes of
bands. These should be prepared according to the manufacturer’s in-
structions.
3. Heat the samples in a water bath or heating block for 2 minutes to de-
nature the samples.
4. Centrifuge the samples in a micro-centrifuge for 20 seconds at 12,000
rpm. The protein samples are now ready to load.
Loading the samples
1. If desired, fit the cooling pack(s) into the end of the tank. These should
be pre-frozen and fitted with the longest side positioned sideways with
the end(s) of the tank and pressed into the recess. Or these can be fit-
ted down the front of the tank.
NEVER FIT THESE UNDERNEATH THE MODULE IN THE BOTTOM OF THE TANK AS THIS WILL PREVENT THE FLOW OF
CURRENT THROUGH THE GEL AND CAUSE SLOW RUNS AND OVER-HEATING.
2. Note one pack is supplied as standard. Additional packs can be pur-
chased.
3. Transfer the Inner gel module containing cast gels into the main tank in
the correct orientation as indicated - +ve on the module aligned with
+ve on the tank, -ve on the module aligned with –ve on the tank.
4. Fill the outer tank with 1X reservoir buffer.
5. Load the samples into the wells using a pipette tip taking care not to
damage the wells or induce any air bubbles.
6. Fill any unused wells with 1X sample buffer.
7. It is a good idea to note the orientation and order the samples were
loaded in. This can be done by noting which samples were loaded ad-
jacent to each electrode.

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Buffer Volumes
Buffer Volume
VS20WAVE
VS10W
Minimum –Inner tank is filled to above the
wells. Outer Tank is filled to just flood the bot-
tom of the glass plates. Cooling potential is
at a minimum which may affect resolution.
Total Inner Buffer
Chamber: 640ml
Outer Tank: 1L
Total Inner Buffer Cham-
ber: 300ml
Outer Tank: 500ml
Maximum –Inner tank is filled to above the
wells. Outer Tank is filled to the maximum fill
line. Cooling is high offering good resolution
of samples.
This may be further enhanced by using the
cooling coil
Total Inner Buffer
Chamber: 640ml
Outer Tank for 2 gels:
5.4L
Outer Tank for 4 gels:
4.8L
Total Inner Buffer Cham-
ber: 300ml
Outer Tank for 2 gels:
2.8L
Outer Tank for 4 gels:
2.3L
Gel Running
1. Fit the lid and connect to a power supply.
2. Consult the below table for details on recommended power supply volt-
age settings.
3. Turn the power supply off when the loading dye reaches the bottom of
the gel, sooner if your proteins are below 4Kd in size.
4. Remove the gel running module, first emptying the inner buffer into the
main tank. Buffer can be re-used but this may affect run quality if contin-
ued.
5. Unscrew the glass plates and gently pry apart the glass plates. The gel
will usually stick to one of the plates and can be removed by first soaking
in buffer and then gently lifting with a spatula.
6. The gel is now ready to be stained with Coomassie or silver stain or the
proteins in the gel can be transferred to a membrane by electroblotting
for specific band identification and further analysis.

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Recommended Voltages for 1mm thick, 12%
gels.
VS20WAVE
VS10WD
2-4 gels
Constant Voltage
250V
6 Hours
Constant Voltage
125V
1 Hour
We recommend that for any new sample, a time course experiment is per-
formed to obtain the optimal running conditions.
Connecting a recirculating chiller
Vertical PAGE may sometimes result in high currents. High current may cause
the build-up of heat which can affect sample migration and gel resolution as
a result. To counteract the adverse effects of heat generation, the manu-
facturer recommends using an effective chiller unit, such as the CSL-CHILLER,
pre-set to 4°C.
1. For active temperature regulation attach two short lengths of hose from
the inlet and outlet ports of the chiller unit to the respective outlet and
inlet connectors of the cooling coil sitting within the WAVE outer tank.
2. Ensure that the flow rate of the chiller unit is set at an acceptable value
by first testing with the cooling coil outside of the electrophoresis tank.
The unit is now ready to be used with active cooling.
Solutions (For SDS-PAGE)
Stock 30% Acrylamide Gel Solution:
30.0 g acrylamide
0.8 g methylene bisacrylamide
Distilled Water to 100ml
Stock 4 X Resolving Gel Tris (1.5 M Tris.HCl pH8.8, 0.4 % SDS)
To 110ml Distilled Water add 36.4 g of Tris base
Add 8ml of 10 % SDS
Adjust pH to 8.8 with 1N HCl
Adjust the final volume to 200ml with Distilled Water.
Stock 4 X Stacking Tris (0.5 M Tris.HCL pH6.8, 0.4 % SDS)
To 110ml Distilled Water add 12.12 g of Tris base

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Add 8ml of 10 % SDS
Adjust pH to 6.8 with 1N HCl
Add Distilled Water to a final volume of 200ml
Stock 4 X Tris-glycine tank buffer - SDS
36 g Tris base
172.8 g glycine
Distilled Water to 3 L
1 x Tris-glycine tank buffer - SDS
750ml of 4 X Tris-glycine reservoir buffer - SDS
30ml of 10 % SDS
Distilled Water to 3L
10 % AP (ammonium persulphate solution)
0.1 g ammonium persulphate
1ml Distilled Water
TEMED
Stock 4 X Sample Buffer
4ml glycerol
2ml 2-mercaptoethanol
1.2 g SDS
5ml 4 X Stacking Tris
0.03 g Bromophenol blue
Aliquot into 1.5ml micro centrifuge tubes. Store at -20°C.
References
1. Sambrook, Fritsch, and Maniatis, Molecular Cloning A Laboratory Man-
ual, Second Edition, Cold Spring Harbor Laboratory Press, 1989.
2. Current Protocols in Molecular Biology, Greene Publishing Associates
and Wiley-Interscience, 1989.

27/11/2020 Page 19
Troubleshooting
Problem: Sample Prepara-
tion
Cause
Solution
Laemmil sample buffer turns yel-
low
Sample buffer too acidic
Add Tris base until buffer turns blue again.
Sample very viscous
High DNA or carbohydrate con-
tent
Fragment DNA with ultrasonic waves during
cell lysis and protein solubilization.
Add endonucleases (for each benzonases).
Precipitate protein with TCA/acetone to di-
minish carbohydrate content.
Problem: Gel casting and
sample loading
Cause
Solution
Poor well formation
Incorrect catalyst used
Monomer solution not degassed
(oxygen inhibits polymerization)
Prepare Fresh catalyst solution.
Increase catalyst concentration of stacking
gel to 0.06% APS and 0.12% TEMED.
Degas monomer solution immediately prior to
casting stacking gel.
Webbing; excess acrylamide be-
hind the comb
Incorrect catalyst concentration
Prepare fresh catalyst solution.
Increase catalyst concentration of stacking
gel to 0.06% APS and 0.12% TEMED.
Gel does not polymerize
Too little or too much APS or
TEMED
Failure to degas
Temperature too low
Poor quality acrylamide or bis
Old APS
Use 0.0.05% APS and 0.05% TEMED.
Degas monomer solutions 10-15min.
Cast at room temperature, warming glass
plates if necessary.
Use electrophoreses-grade reagents
Prepare fresh APS.
Swirls in the gel
Excess catalysts; polymerization
time < 10min
Gel inhibition; polymerization
time >2hr
Reduce APS and TEMED by 25% each.
Increase APS and TEMED by 50%; degas.
Gel feels soft
Low %T
Poor quality acrylamide or bis
Too little cross-linker
Use different %T.
Use electrophoresis- grade reagents.
Use correct %C.
Gel turns white
Bis concentration too high
Check solutions or weights.
Gel brittle
Cross-linker too high
Use correct % cross-linker
Sample floats out of the well
Sample is not dense enough
Pipetting, loading error
Induce 10% glycerol in sample to make it
denser than surrounding buffer.
Slowly pipet sample into well. Do not squirt
sample quickly into well as it may bounce off
bottom or sides and flow into next well. Do
not pipet tip from well before last of sample
has left the tip.
Problem: Electrophoresis
Cause
Solution
Current zero or less than ex-
pected and samples do not mi-
grate into gel
Tape at the bottom of precast
gel cassette not removed
Insufficient buffer in inner buffer
chamber
Remove tape.
Fill buffer chamber with running buffer.

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Insufficient buffer in outer buffer
chamber
Electrical disconnection
Fill inner and outer buffer chambers to ensure
wells are completely covered.
Check electrodes and connections.
Gels run faster than expected
Running buffer too concentrated
and gel temperature too high; in-
correct running buffer concen-
tration or type used
Running or reservoir buffer too di-
lute
Voltage too high
Check buffer composition and type.
Check buffer protocol and concentrate if
necessary.
Decrease voltage by 25-50%.
Gels run slower than expected
Incorrect running buffer composi-
tion or type
Excessive salt in sample
Check buffer composition and type.
Desalt sample.
Problem: Total Protein
Staining
Cause
Solution
Bands not visible
No protein in gel
Imaging system malfunctioning
Incorrect imaging parameters
were used
Stain with another method to confirm there is
protein.
Check instrument manual for troubleshooting
or contact imaging instrument manufacturer.
Check Instrument manual.
Poor staining sensitivity
Dirty staining trays
Insufficient stain volume
Insufficient staining time
Reuse of staining solution
Clean staining trays and other equipment
with laboratory glassware cleaner.
Follow recommendations for stain volume
(appropriate to gel size).
Increase staining time.
Repeat staining protocol with fresh staining
solution.
High or uneven background
staining
Staining trays or equipment dirty
Too much time in staining solu-
tion
Reagent impurities
Clean staining trays and other equipment
with laboratory glassware cleaner.
Restrict duration of incubation in staining so-
lutions as recommended in protocol.
Wash gel in water or retrospective destaining
solution for >30min.
Use high-purity water and reagents for stain-
ing.
Speckles or blotches in gel im-
age
Particulate material from rea-
gents, staining tray, dust or
gloves
Clean staining trays thoroughly.
Decrease time that gels and staining solution
are exposed to open air.
Use dust-free gloves and handle gels only by
edges.
Uneven staining
Insufficient shaking during stain-
ing
Agitate gel during staining.
Gel shrinkage
Gel dehydrated
Transfer gel to water for rehydration.
Problem: Evaluation of
Separation
Cause
Solution
Diffuse or broad bands
Poor quality acrylamide or bis-
acrylamide incomplete polymeri-
zation
Old SDS or sample buffer
Gel temperature too high
Use electrophoresis-grade reagents.
Check polymerization conditions.
Prepare fresh solutions.
Use external cooling during run or run out a
lower voltage.
Bands ‘smile’ across gel, band
pattern curves upward at both
sides of gel
Excess heating of gel; centre of
gel runs hotter than either end
Check buffer composition; buffer not mixed
well, or buffer in upper chamber too concen-
trated.