Consort EVS1000 Series User manual

EVS1000 BLOT series
MANUAL
Rev. 44/2019

Contents
Contents .....................................................................................................................................................................................................................b
Safety Information ................................................................................................................................................................................................ 1
Precaution........................................................................................................................................................................................................... 1
General care and maintenance..................................................................................................................................................................... 1
Environmental conditions.................................................................................................................................................................................. 2
Fitting electrode cables ..................................................................................................................................................................................... 2
Electroblotting......................................................................................................................................................................................................... 3
Setting up the blot sandwich...................................................................................................................................................................... 3
Blot run conditions .......................................................................................................................................................................................... 3
Buffer solutions for blotting........................................................................................................................................................................ 4
Protein (Western) blotting buffers ......................................................................................................................................................... 5
Nucleic acid transfer buffers..................................................................................................................................................................... 5
Transfer buffers ............................................................................................................................................................................................... 5
Certificate ................................................................................................................................................................................................................. 6
Warranty................................................................................................................................................................................................................... 7

EVS1000 Blot units
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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 inner module should be thoroughly rinsed with warm water or distilled water to prevent buildup
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.

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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”.
Fitting electrode cables
Note: Before setting up the tank please ensure that it has been properly cleaned and dried.
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 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.

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Electroblotting
Setting up the blot sandwich
Each blot sandwich should be set up as follows:
•Cassette clamp [-ve] (black) side placed in a tray or other suitable surface.
•Pre-soaked fibre pad (note two can be used with thin gels).
•Two pieces of thick filter paper, about 2...3 mm thick, pre-soaked in buffer.
•Gel.
•Transfer membrane. Usually this requires pre-soaking but consult the manufacturer’s
instructions for the type of membrane you are using. This should be smoothed so that no air
bubbles have been trapped.
•Two pieces of thick filter paper, about 2...3 mm thick, pre-soaked in buffer.
•Pre-soaked fibre pad (note two can be used with thin gels).
•Cassette clamp [+ve] (red) side slotted into the groove in the bottom of the black cassette.
Close the hinge carefully so as to not disturb the sandwich.
Fill the tank with buffer solution up to the maximum fill line indicated on the side of each unit.
Improved transfer can usually be obtained by using chilled buffer.
Buffer Volume
EVS1100
EVS1200
EV1300
One cassette
1380 ml
2800 ml
5600 ml
Two cassettes
1290 ml
2620 ml
5240 ml
Three cassettes
1200 ml
2440 ml
4880 ml
Four cassettes
1110 ml
2260 ml
4520 ml
Each cooling pack will take the place of 100 ml of buffer for EVS1100 and 500 ml of buffer for
EVS1200 and EVS1300.
Blot run conditions
Insert the cassettes into the slots in the module with the black side of each adjacent to the negative
electrode. It is a good idea to note the orientation and order the blot sandwiches were loaded in. This
can be done by noting which samples were loaded adjacent to each electrode.
Use of a magnetic stirring bar and plate is recommended to mix the buffer to give consistency of
transfer. A 4 mm diameter stirring bar should be placed underneath the module, in the centre of the
tank. The cooling pack provided, pre-frozen, can be inserted at the side or front of the tank for
extended blots. Additional cooling packs can be purchased as accessories to further aid cooling.
Insert the module, fit the lid and connect to a power supply.
Consult below table for details on recommended power supply voltage settings and blot times. Please
note voltage and current will vary according to the amount of cassettes, type and temperature of

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buffer and thickness and percentage of gel. This will also affect quality of transfer so time course of
transfer should be performed for your particular samples and conditions.
When the blot time is completed, turn the power supply off.
Remove the cassettes from the main tank. Buffer can be reused but this may affect run quality if
continued.
Lift the hinge of each cassette and gently pry apart the blot sandwich and remove the membrane
from the gel.
The membrane is now ready to be probed.
Duration of blotting
EVS1100
EVS1200
EV1300
One hour
100 V
400 mA
100 V
400 mA
100 V
400 mA
Three hours
50 V
200 mA
50 V
200 mA
50 V
200 mA
Buffer solutions for blotting
Do not adjust the pH when making these buffers as this will cause blot over-heating. The pH will vary
according to the freshness of the reagents used.
Towbin Buffer
•25 mM Tris,
•192 mM glycine,
•20 % methanol pH 8.3
Towbin Buffer SDS
•25 mM Tris
•192 mM glycine
•20 % methanol pH 8.3
•0.05-0.1 % (w/v) SDS
Bjerrum and Schafer-Nielsen Buffer
•48 mM Tris
•39 mM glycine
•20 % methanol pH 9.2
Dunn Buffer
•10 mM NaHCO3
•3 mM NaCO3
•20 % methanol pH 9.9

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Protein (Western) blotting buffers
Transfer buffers must be made accurately using high grade reagents. Do not adjust the pH
with acid or base as this will affect the properties of the buffer. pH will vary according to the
purity of the reagents used.
Triple buffer system
For high efficiency transfer of Protein from acrylamide gels
•Anode 1 Buffer:
0.3 M Tris base, 20% MeOH, pH 10.4:
soak 4 standard grade filter paper sheets
•Anode 2 Buffer:
0.025 M Tris base, 20% MeOH pH 10.4:
soak 2 standard grade filter paper sheets
•Cathode Buffer:
0.025 M Tris base, 0.04 M Caproic Acid, 20% MeOH pH 9.4:
soak 6 standard grade filter paper sheets
DNA (Southern) Blotting Buffer
For high efficiency transfer of DNA from agarose gels.
•Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in:
o50 x 1 M ethanolamine-glycine buffer, pH 11
RNA (Northern) Blotting Buffer
For high efficiency transfer of RNA from agarose gels.
•Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in:
o50 x 0.2 M morpholinopropanesulfonic acid (MOPS)
o50 mM sodium acetate
o5 mM EDTA pH 7.0
Nucleic acid transfer buffers
Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in:
•1 x TAE 40 mM Tris (pH 7.6), 20 mM acetic acid, 1 mM EDTA
•50 x (1l) dissolve in 750 ml distilled water:
o242 g Tris base (FW = 121)
o57.1 ml glacial acetic acid
o100 ml 0.5 M EDTA (pH 8.0)
•Fill to 1 litre with distilled water.
Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in:
•1 x TBE 89 mM Tris (pH 7.6), 89 mM boric acid, 2 mM EDTA
•10 x (1l) dissolve in 750 ml distilled water:
o108 g Tris base (FW = 121)
o55 g boric acid (FW = 61.8)
o40 ml 0.5 M EDTA (pH 8.0)
•Fill to 1 litre with distilled water.
Transfer buffers
Towbin Buffer with 20% methanol
Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in:
•0.025 M Tris base
•0.192 M glycine
•20% methanol
•pH 8.3

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Certificate

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Warranty
The Consort 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, 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!

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EVS1000 Blot units
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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 : sup[email protected]
Information : [email protected]