Consort EHS1000 Series User manual

EHS1000 series
MANUAL
Rev. 44/2019
Gel Electrophoresis
Horizontal units

Models
Gel
Width
(cm)
Length
(cm)
Sample
Capacity
Gel Volume
5mm thickness
(ml)
EHS1050
10
8
40
40
EVS1100
7
7/10
64
25/35
EHS1200
10
7/10
100
35/50
EHS1300
15
7/10/15
210
53/75/113
EHS1400
20
10/20/25
550
100/200/250
EHS1500
26
16/24/32
672
308/312/416

Contents
Models......................................................................................................................................................................................................................... II
Contents .....................................................................................................................................................................................................................c
Safety Information ................................................................................................................................................................................................ 1
Precaution........................................................................................................................................................................................................... 1
General care and maintenance..................................................................................................................................................................... 1
Operating instructions........................................................................................................................................................................................ 2
Environmental conditions............................................................................................................................................................................. 2
Setting up the horizontal gel tank............................................................................................................................................................ 2
Gel preparation................................................................................................................................................................................................. 3
Gel pouring with model EHS1050 .......................................................................................................................................................... 3
Gel pouring with casting dams.................................................................................................................................................................. 4
Gel pouring using the flexicaster.............................................................................................................................................................. 4
Gel pouring using tape method ................................................................................................................................................................ 5
Running the gel ................................................................................................................................................................................................. 5
Gel staining and viewing ............................................................................................................................................................................... 5
Solutions.................................................................................................................................................................................................................... 6
Troubleshooting..................................................................................................................................................................................................... 7
Equipment problem ........................................................................................................................................................................................ 7
Electrophoresis problem.............................................................................................................................................................................. 7
Certificate ................................................................................................................................................................................................................. 9
CE............................................................................................................................................................................................................................. 9
Warranty................................................................................................................................................................................................................10

Horizontal Gel Electrophoresis units
1
Safety Information
Precaution
•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 position.
•The unit should not be used if there is any sign of damage to the external tank or lid.
•Always isolate electrophoresis units from their power supply before removing the safety cover.
Isolate the power supply from the mains first then disconnect the leads.
•Do not exceed the maximum operating voltage or current.
•Do not operate the electrophoresis units in metal trays.
•Acrylamide is a volatile, cumulative neurotoxin and suspected carcinogen. Wear effective protective
clothing and follow recommended handling and disposal procedures.
•Polymerised gels contain some unpolymerised monomer. Handle with gloves only. Following the
replacement of a platinum electrode have the unit inspected and approved by your safety officer
prior to use.
•Do not fill the unit with running buffer above the maximum fill lines.
•Do not move the unit when it is running.
•Caution: during electrophoresis very low quantities of various gases are produced at the electrodes.
The type of gas produced depends on the composition of the buffer employed. To disperse these
gases make sure that the apparatus is run in a well ventilated area.
General care and maintenance
•Units are best cleaned using warm water and a mild detergent. Water at temperatures above
60°C can cause damage to the unit and components. The tank should be thoroughly rinsed with
warm water or distilled water to prevent build up of salts but care should be taken not to damage the
enclosed electrode and vigorous cleaning is not necessary or advised. Air drying is preferably before
use.
•The units should only be cleaned with the following: warm water with a mild concentration of soap or
other mild detergent (compatible detergents include dish washing liquid, hexane and aliphatic
hydrocarbons). The units should not be left to in detergents for more than 30 minutes.
•The units should never come into contact with the following cleaning agents, these will cause
irreversible and accumulative damage: acetone, phenol, chloroform, carbon tetrachloride,
methanol, ethanol, isopropyl alcohol alkalis.
•In case of Rnase Decontamination clean the units with a mild detergent as described above. Wash
with 3 % hydrogen peroxide (H2O2) for 10 minutes. Rinsed with 0.1 % DEPC- (diethyl pyrocarbonate)
treated distilled water (Caution: DEPC is a suspected carcinogen. Always take the necessary
precautions when using.) RNaseZAP™ (Ambion) can also be used. Please consult the instructions for
use with acrylic gel tanks.

Horizontal Gel Electrophoresis units
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Operating instructions
Environmental conditions
•This apparatus is intended for indoor use only.
•This apparatus can be operated safely at an altitude up to 2000 m.
•The normal operating temperature range is between 4°C and 65°C.
•Maximum relative humidity 80 % for temperatures up to 31°C decreasing linearly to 50 % relative
humidity at 40°C.
•The 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 horizontal gel tank
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.
Fitting loading guides
These can be fitted to enhance visibility of the wells if desired. They can be fitted to the white vinyl platform
sheet or to the unit itself.
Seat the tray in the unit and note the position of the comb grooves. The samples run black to red but
the trays can be used frontward or backwards so ensure that the comb grooves closest to the black
electrode are marked.
Remove the tray.
Peel the back off of the loading guide and carefully apply the loading guide directly to the gel platform.
The unit is now ready to be used.

Horizontal Gel Electrophoresis units
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Gel preparation
Model
Gel Width (cm)
Length (cm)
Gel Volume (ml)
EHS1050
10
8
40
EVS1100
7
7
10
25
35
EHS1200
10
7
10
35
50
EHS1300
15
7
10
15
53
75
113
EHS1400
20
10
20
25
100
200
250
EHS1500
26
16
24
32
308
312
416
This table shows the volume of agarose solution required to make the desired agarose gel (5 mm
thick) for each unit tray size. For a standard 0.7 % agarose gel, add 0.7 g of agarose to 100 ml of 1x
TAE or TBE solution. The same 1x solution should be used in the tank buffer solution.
Add the agarose powder to a conical flask.
Add the appropriate amount of 1 x TAE or TBE solution from the table above. To prevent evaporation
during the dissolving steps below, the conical flask should be covered with parafilm.
Dissolve the agarose powder by heating the agarose either on a magnetic hot plate with stirring bar
or in a microwave oven. If using the microwave method, the microwave should be set at around a
400 W or medium setting and the flask swirled every minute. The solution should be heated until all
crystals are dissolved. This is best viewed against a light background. Crystals appear as translucent
crystals. These will interfere with sample migration if not completely dissolved.
The gel must be cooled to between 50°C and 60°C degrees before pouring.
Gel pouring with model EHS1050
Fit the casting gates into the grooves in the unit which are 10 mm from the platinum wire. Ensure
that these are seated as deeply as possible. This will ensure that a good seal is formed and that
there is no possibility of gel leakage. A small amount of vaseline or sealing grease on the bottom and
side edges of the gates will give further leakage protection.
Place the comb(s) in the grooves. Each tray has more than one comb grove so that multiple combs
can be used. Using multiple combs increases sample number available per gel but decreases run
length and care must be taken to ensure that samples from the first wells do not migrate into the
lanes of the second comb wells.
Pour in the agarose carefully so as not to generate bubbles. Any bubbles that do occur can be
smoothed to the edge of the gel and dispersed using a pipette tip.
Allow the agarose to set, ensuring that the gel remains undisturbed.
Carefully remove the gel casting gates and comb and transfer the gel including tray to the main tank.

Horizontal Gel Electrophoresis units
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Gel pouring with casting dams
Fit the casting dams over each end of the tray and place onto a level surface. The dams should be
fitted so that there is no gap between the sides of the tray and the groove in the dams. This will
ensure that there is no possibility of gel leakage.
Place the comb(s) in the grooves. Each tray has more than one comb grove so that multiple combs
can be used. Using multiple combs increases sample number available per gel but decreases run
length and care must be taken to ensure that samples from the first wells do not migrate into the
lanes of the second comb wells.
Pour in the agarose carefully so as not to generate bubbles. Any bubbles that do occur can be
smoothed to the edge of the gel and dispersed using a pipette tip.
Allow the agarose to set, ensuring that the gel remains undisturbed.
Carefully remove the gel casting gates and comb and transfer the gel including tray to the main tank.
Gel pouring using the flexicaster
Level the flexicaster base by adjusting the feet so that the bubble is exactly central.
Insert the desired length tray into the flexicaster such that one end of the tray is pushed up and seals
against the silicone mat of the permanent end of the flexicaster.
Position the movable end of the flexicaster so that the silicone mat is pushed against the other end of
the tray.
Turn the cam so that the silicone mat tightly seals against the side of the tray. Pour in the agarose
carefully so as not to generate bubbles. Any bubbles that do occur can be smoothed to the edge of
the gel and dispersed using a pipette tip.
Allow the agarose to set, ensuring that the gel remains undisturbed.
Carefully remove the gel casting gates and comb and transfer the gel including tray to the main tank.

Horizontal Gel Electrophoresis units
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Gel pouring using tape method
1. Autoclave or plastic backed general tape should be used. A length 5 cm longer than the width of
each end of the tray should be cut. One length should be placed over one end of the tray folded
and the edges sealed securely. Repeat for the other end and place onto a level surface for gel
pouring.
2. Place the comb(s) in the grooves. Each tray has more than one comb grove so that multiple
combs can be used. Using multiple combs increases sample number available per gel but
decreases run length and care must be taken to ensure that samples from the first wells do not
migrate into the lanes of the second comb wells.
3. Pour in the agarose carefully so as not to generate bubbles. Any bubbles that do occur can be
smoothed to the edge of the gel and dispersed using a pipette tip.
4. Allow the agarose to set, ensuring that the gel remains undisturbed.
5. Carefully remove the gel casting gates and comb and transfer the gel including tray to the main
tank.
Running the gel
Mix the sample to be loaded with sample buffer (see solutions for common sample buffers). Usually 3
µl of sample buffer is adequate but less may be used with sample volumes of less than 10 µl.
Fill the unit with buffer until the gel is just flooded with buffer. This will give the fastest resolution
times. For enhanced quality of resolution of sample, fill the unit to 5 mm above the gel.
Load the samples into the wells using pipettes. Multi-channel pipettes can be used for loading
samples with MultiPipette compatible combs, see listing in accessories for identification of these.
Carefully place the lid on the tank and connect to a power supply.
Typically, gels are run at between 90 and 150 V. However, maximum voltage is indicated on the
serial badge of each unit. It should be noted that a higher voltage generally gives faster but poorer
quality sample resolution.
Gel staining and viewing
All trays and the EHS1050 unit allow staining to be performed without removing the gel from the tray if
this is preferred.
1. Transfer the gel to a vessel containing the appropriate volume of 0.5 µg/ml ethidium bromide
stain for 15...30 minutes, see solutions for stock stain concentration and adjust to the volume
used accordingly. The entire gel should be covered. (Caution: ethidium bromide is a suspected
carcinogen and the necessary safety precautions should be undertaken).
2. De-stain the gel for 10–30 minutes in distilled water again ensuring the gel is completely
immersed.
3. Rinse the gel twice for a couple of seconds with distilled water.
4. Transfer the gel to a UV Transilluminator.
5. The samples will often appear as brighter, clearer bands when photographed or viewed using a
gel documentation system. However, if the gel bands are too faint then the staining procedure
should be adjusted so that there is less de-staining. If there is too much background, then the
staining procedure should be adjusted so that there is more de-staining.

Horizontal Gel Electrophoresis units
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Solutions
1x TAE 40 mM tris (pH 7.6), 20 mM acetic acid, 1 mM EDTA
50 x (1l), dissolve in 750 ml distilled water:
242 g tris base (FW = 121)
57.1 ml glacial acetic acid
100 ml 0.5 M EDTA (pH 8.0).
Fill to 1 litre with distilled water.
1x TBE 89 mM tris (pH 7.6), 89 mM boric acid, 2 mM EDTA
10 x (1l), dissolve in 750 ml distilled water:
108 g tris base (FW = 121)
55 g boric acid (FW = 61.8)
40 ml 0.5 M EDTA (pH 8.0)
Fill to 1 litre with distilled water.
Sample Loading Dye
10x sample buffer stock consists of 50% glycerol, 0.25% bromophenol blue, and 0.25% xylene cyanole FF in
1x TAE buffer. Only 1–10 ml of the 10x loading dye should be prepared.
Ethidium Bromide Solution
Add 10 mg of Ethidium Bromide to 1 ml distilled water.

Horizontal Gel Electrophoresis units
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Troubleshooting
Equipment problem
Bubbles do not appear on the electrodes:
•Ensure that the power supply and the whole electric assembly is operating properly.
Melted agarose leaks when casting:
•Ensure the agarose is not too hot when poured.
•Ensure that the sealing surfaces of the running tray and the gel casting gates are clean.
•Ensure that the ends of the running tray are flat and free of nicks.
Electrophoresis problem
Sample well deformed:
•Allow the gel to set for a minimum of 30 minutes.
•Leave comb in position until gel returns to room temperature before removing.
•Remove the comb both slowly and at a slight angle to prevent gel from breaking.
•Avoid damaging the well with the pipette when loading the sample. Aim for the centre of the well and
avoid damaging the bottom of the well with the pipette tip.
Samples leak underneath the gel upon loading:
•The bottom of the wells were torn when the comb was removed. To avoid this tearing, carefully
wiggle the comb to free the teeth from the gel.
Distorted sample wells:
•Incomplete polymerisation produces poorly defined wells.
•De-gas gel solution prior to casting and increase ammonium persulphate and TEMED oncentrations.
Samples do not run straight:
•Comb may be warped and should be replaced.
•Running tray may be warped and should be replaced. Reduce the voltage.
•Choose a buffer with suitable ionic strength and buffering capacity.
“Smiling” along one edge of the gel:
•Gel was not level when cast or run. Use a gel levelling table to ensure that the apparatus is level prior
to gel casting and electrophoresis.
Bromophenol blue dye turns yellow:
•Check pH of buffer during electrophoresis. (pH change).
•Ensure Tris base and not Tris-HCl was used.
•Mix the buffer periodically during electrophoresis. Connect a pump to circulate the buffer.
Double-banded pattern
•Ensure the comb is vertical during casting so that the well shape is not distorted.
•Decrease the buffer level to 1 mm above the top of the gel. This will reduce the temperature
gradient through the gel.
•Increase concentration of the sample and use a thin (2 to 3 mm) gel with a thin (1 mm) comb.
"Tailed" bands (excessive fluorescence appearing above the band)
•Reduce DNA in the sample.
•Reduce the protein and/or glycerol in the sample.
• Poor band resolution
•Add ficoll, glycerol, or sucrose to the sample loading buffer to ensure that the sample layers on the
bottom of the well. Ensure sample is completely dissolved.
•Reduce voltage, sample concentration, or sample volume.
•Ensure there is at least 1 mm of gel below the bottom of the comb to prevent samples from leaking
out the bottom of the well.

Horizontal Gel Electrophoresis units
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•Reduce salt concentration of the sample. High salt concentrations can cause "pinched" lanes,
smeared lanes, arched dye front and slow migration.
•Check enzyme activity as it may require longer digestion or different restriction buffer.
•Prepare fresh sample if nuclease contamination is suspected.
•Choose agarose with low endosmosis value.
Gel melts or softens near sample wells.
•Caused by a combination of pH drift and high temperature. Circulate or remix buffer periodically or
reduce the voltage.

Horizontal Gel Electrophoresis units
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Certificate
CE

Horizontal Gel Electrophoresis units
10
Warranty
The Consort Horizontal Electrophoresis units have a warranty against manufacturing and material faults of
twelve months from date of customer receipt.
If any defects occur during this warranty period, Consort will repair or replace the defective parts free of
charge. This warranty does not cover defects occurring by accident or misuse or defects caused by improper
operation.
Units where repair or modification has been performed by anyone other than Consort or an appointed
distributor or representative are no longer under warranty from the time the unit was modified.
Units which have accessories or repaired parts not supplied by Consort or its associated distributors have
invalidated warranty.
Consort cannot repair or replace free of charge units where improper solutions or chemicals have been
used. For a list of these please see the Care and Maintenance subsection.
Consort products are for research use only. Consort is not liable for consequential damages arising out of
the use or handling of its products.
A return authorisation must be obtained from Consort before returning any product for warranty repair on a
freight prepaid basis. If a problem does occur then please contact your supplier or Consort:
Consort bvba
Hertenstraat 56/9
2300 Turnhout
Belgium
Tel: [phone]
Email: [email protected]
Record the following for your records:
Model __________________________
Date of Delivery __________________
Warranty Period __________________
Serial No. _______________________
Invoice No. ______________________
Purchase Order No. _______________
A return authorisation must be obtained from CONSORT before returning any product for warranty repair on
a freight prepaid basis!

Horizontal Gel Electrophoresis units
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Horizontal Gel Electrophoresis units
12
Consort bvba
Hertenstraat 56 unit 9
2300 Turnhout
Belgium
Tel : (+32) (0)14 41 12 79
Fax : (+32) (0)14 42 91 79
Sales : [e-mail]
Support : [e-mail]
Information : [email protected]