Eco-Line user manual (English)
Eco-Line
Vertical Polyacrylamide Gel Electrophoresis
&
Tank Blotting Apparatus
Instruction Manual
Model
Order No
Eco-Mini E / System E
Eco-Mini EB / System EB
Eco-Mini EBC / System EBC
Eco-Maxi EB / System EB
Eco-Maxi EBC / System EBC
Tankblot Eco-Mini C
Tankblot Eco-Mini
Tankblot Eco-Maxi C
Tankblot Eco-Maxi
017-101 / 017-100
017-103 / 017-102
017-105 / 017-104
017-400 / 017-401
017-402 / 017-403
018-100
018-101
018-400
018-401
Please read these instructions carefully
before using this apparatus!
Biometra GmbH
Rudolf-Wissell-Str. 30
D-37079 Goettingen
Tel:
+49 (0)5 51 50 68 6-0
Fax:
+49 (0)5 51 50 68 6-66
e-mail: [email protected]
internet: http://www.biometra.de
Service Department
Rudolf-Wissell-Str. 14-16
D-37079 Goettingen
Tel.: +49 (0)5 51 50 68 6-10 or -12
Fax.: +49 (0)5 51 50 68 6-11
e-mail: [email protected]
This document describes the state at the time of publishing. It needs not necessarily agree
with future versions.
Subject to change!
Eco_Line Manual, 2014_06
1
Contents:
1
2
3
4
5
6
Introduction.......................................................................................................... 3
1.1
Field of Application .......................................................................................... 3
1.2
Specifications .................................................................................................. 5
1.3
Legal Notes ..................................................................................................... 7
1.4
Meaning of the Instructions ............................................................................. 7
Safety and Warning Notices ............................................................................... 8
2.1
Definition of Symbols ....................................................................................... 8
2.2
General Safety Instructions ............................................................................. 8
Setting-Up Operation ........................................................................................ 11
3.1
Scope of Delivery .......................................................................................... 11
3.2
Unpack and Check ........................................................................................ 15
3.3
Installation Conditions ................................................................................... 15
3.4
Connecting Conditions .................................................................................. 15
Components....................................................................................................... 16
4.1
Systems Overview ......................................................................................... 16
4.2
Compatibility Overview .................................................................................. 18
Electrophoresis ................................................................................................. 19
5.1
Gel Cassette Assembly ................................................................................. 19
5.2
Gel Casting .................................................................................................... 21
5.3
System Assembly and Sample Loading ........................................................ 22
5.4
Electrophoresis .............................................................................................. 23
5.5
System Reassembly and Gel Removal ......................................................... 23
Tankblotting ....................................................................................................... 24
6.1
Assembling the Gel Sandwich ....................................................................... 24
6.2
Setting up the Buffer Tank (EB or EBC) ........................................................ 25
6.3
Transfer of more than one Gel ...................................................................... 25
6.4
Transfer Times and Conditions ..................................................................... 26
6.5
System Reassembly and Blot Removal ......................................................... 26
Eco Line Manual, Vers. 2014_06
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7
Buffers and Staining/Destaining Solutions ..................................................... 27
7.1
Gel Solutions: ................................................................................................ 27
7.2
Electrophoresis Buffer Solutions: .................................................................. 30
7.3
Blotting Buffer Solutions: ............................................................................... 31
7.4
Staining and Destaining Solutions: ................................................................ 32
8
Trouble Shooting ............................................................................................... 33
8.1
Electrophoresis .............................................................................................. 33
8.2
Tankblotting ................................................................................................... 36
9
Product Information .......................................................................................... 38
10
Maintenance and Repair ................................................................................ 44
10.1
Cleaning and Maintenance ......................................................................... 44
10.2
Servicing ..................................................................................................... 44
10.3
Replacement of Spare Parts ...................................................................... 44
10.4
Other Accessories ...................................................................................... 44
11
Service ............................................................................................................. 45
11.1
Instructions for Return Shipment ................................................................ 45
12
Equipment Decontamination Certificate....................................................... 46
13
Note for Disposal of Electric/Electronic Waste ............................................ 48
14
EC – Declaration of Conformity ..................................................................... 49
15
Warranty .......................................................................................................... 50
Eco_Line Manual, 2014_06
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1
Introduction
1.1
Field of Application
The Biometra Eco-Line is a modular, tank-style apparatus designed for the
separation of proteins and nucleic acids (DNA/RNA) in vertical polyacrylamide
(PAGE) gels.
Interchangeable electrode modules allow electrophoresis and blotting using the
same buffer tanks. Using the Electrophoresis Module (Gel Module) the Eco-Line runs
up to 4 gels (Eco-Mini) or up to two gels (Eco-Maxi). The module accepts hand cast
gels as well as pre-cast gels (Eco-Mini only). Using the Blot Module the apparatus
allows blotting of one to two gels (Eco-Maxi) or one to four gels (Eco-Mini)
simultaneously in individual cassettes.
Eco-Mini system E
(017-100)
Eco-Mini system EB
(017-102)
Eco-Mini system EBC
(017-104)
Tankblot Eco-Mini C
(018-100)
Tankblot Eco-Mini
(018-101)
1
2

Components included in the system
Buffer tank
Cooling
Blotting
Electrophoresis
Suitability
E



1

EBC
2


EBC
2

1
EB
EB
furthermore
Buffer tank (see left), bigfoot lid,
Electrophoresis Module, 2 sets of glass plates
with fixed 1 mm spacers, 1 dummy plate,
2 combs (10 wells), casting stand and manual
Buffer tank (see left), bigfoot lid,
Blot Module, 4 blotting cassettes (colour-coded),
8 fiber pads and manual
with addition of components required for blotting (see "furthermore" for Tankblot Eco-Mini systems)
with addition of components required for electrophoresis (see "furthermore" for Eco-Mini systems)
Eco Line Manual, Vers. 2014_06
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1
2
1

1

EBC
2


EBC
2

Buffer tank

Cooling
Blotting
Eco-Maxi system EB
(017-400)
Eco-Maxi system EBC
(017-402)
Tankblot Eco-Maxi C
(018-400)
Tankblot Eco-Maxi
(018-401)
Components included in the system
Electrophoresis
Suitability
EB
EB
furthermore
Buffer tank (see left), bigfoot lid,
Electrophoresis Module, 2 sets of glass plates
with fixed 1 mm spacers, 2 combs (12 wells),
casting stand and manual
Buffer tank (see left), bigfoot lid,
Blot Module, 2 blotting cassettes (colour-coded),
4 fiber pads and manual
with addition of components required for blotting (see "furthermore" for Tankblot Eco-Maxi systems)
with addition of components required for electrophoresis (see "furthermore" for Eco-Maxi systems)
The Biometra Eco- Line is designed for laboratory research only.
Eco_Line Manual, 2014_06
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1.2
Specifications
Construction
Buffer Tank
Bigfoot safety lid
PMMA
PMMA
Blotting Module
Blot cassette
PMMA
PMMA
Electrophoresis (Gel) Module
POM
Casting Stand
PE
Combs
Teflon
Eco-Mini combs:
Thickness
(mm)
0.75
1.0
1.5
(c)
Number of Wells
Volume per Well
(µl)
1 prep.
9 MTP(c)
10
12
15
2 x 25 and 385
30
33
25
18
1 prep.
9 MTP(c)
10
12
15
2 x 35 and 515
43
45
35
25
1 prep.
9 MTP(c)
10
12
15
2 x 55 and 780
65
70
55
37
Microtiter plate compatible / multichannel pipet compatible
Eco-Maxi combs:
Thickness
(mm)
0.75
1.0
1.5
(c)
Number of Wells
Volume per Well
(µl)
1 prep.
12
19 MTP(c)
25
30
2 x 30 and 970
70
32
30
20
1 prep.
12
19 MTP(c)
25
30
2 x 45 and 1,310
95
45
40
29
1 prep.
12
19 MTP(c)
25
30
2 x 75 and 2,340
165
75
70
52
Microtiter plate compatible / multichannel pipet compatible
Eco Line Manual, Vers. 2014_06
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Buffer volume
Eco-Mini
approx. 1,580 ml for electrophoresis
(buffer tank E)
approx. 2,180 ml for electrophoresis
(buffer tank EB and EBC)
Approx. 2,100 ml for blotting
(buffer tank EB and EBC)
Eco-Maxi
approx: 5,400 ml for electrophoresis
(buffer tank EB and EBC)
approx. 6,100 ml for blotting
(buffer tank EB and EBC)
Cooling option
Overall size (W x D x H)
Buffer tank EBC: flow through
Eco-Mini
21.0 x 15.0 x 22.0 cm (buffer tank E)
21.0 x 15.0 x 25.0 cm (buffer tank EB)
21.0 x 15.0 x 26.5 cm (buffer tank EBC)
Eco-Maxi
32.5 x 15.0 x 38.5 cm (buffer tank EB)
32.5 x 15.0 x 40.0 cm (buffer tank EBC)
Gel size (W x L)
Eco-Mini
Eco-Maxi
9.4 x 8.0 cm
19.4 x 18.5 cm
Blotting area (W x L)
Eco-Mini
Eco-Maxi
9.4 x 8.0 cm
22.0 x 19.0 cm
Gel compatibility
Eco-Mini
Eco-Maxi
hand cast and pre-cast gels
hand cast gels
Weight
Eco-Mini
Eco-Maxi
max. 2.5 kg (depending on system)
max. 8 kg (depending on system)
Chemical compatibility
The components are not compatible with
alcohol >10% (e.g. ethanol) or organic solvents
(e.g. acetone, chloroform, toluene, benzene).
Use of organic solvents voids all warranties.
The instrument is designed for operation in closed laboratory rooms at ambient temperature
from +5 °C to +40 °C and maximum relative humidity 80% for temperatures up to +31 °C
decreasing linearly to 50% relative humidity at +40 °C.
Eco_Line Manual, 2014_06
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1.3
Legal Notes
Copyright
All rights reserved. It is not allowed to copy and publish the manual or parts of it in any form
as copies, micro film or other methods without a written authorisation from Biometra.
Biometra is pointing out that applied company and brand names are usually protected trade
marks.
Liability
Biometra is not liable for damages and injuries caused by use not considering these
operation instructions in parts or completely.
1.4
Meaning of the Instructions
Biometra recommends that you first read these instructions carefully. Then assemble and
disassemble the apparatus to become familiar with the system. After these preliminary
steps, you should be ready to transfer a sample.
This operation instruction is part of the product and should be kept over the full life-time of
the instrument. It should also be forwarded to subsequent owners and users. Make sure that
additions and updates are inserted into the operation instructions.
Eco Line Manual, Vers. 2014_06
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2
2.1
Safety and Warning Notices
Definition of Symbols
Symbol
Definition
Caution! Refer to instruction manual!
Danger! High voltage!
Fragile!
Using with direct current (DC).
2.2
General Safety Instructions
Delicate instruments! Handle with care!
Do not operate this equipment if buffer or water leaks from the
instrument, if cracks are present in the body or the safety cover or if the
electrical connection cables are worn or frayed.
Never place the instrument on top of a Power Supply.
Make sure that the on/off switch of the used external Power Supply is
always free accessible.
Use only Biometra safety power cords for connecting the
electrophoresis equipment to the Power Supply.
Eco_Line Manual, 2014_06
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Danger! High voltage!
The current to the cell, provided from the external Power Supply, enters
the unit through the lid assembly, providing a safety interlock to the
user. Current to the cell is broken when the lid is removed.
the
Do not attempt to circumvent this safety interlock, and always turn
Power Supply off before removing the lid, or when working with
the cell in any way.
Power to the instrument is supplied by an external DC voltage Power
Supply. The output of this Power Supply must be isolated from external
ground to issue that the DC voltage output floats with respect to ground.
(All Biometra Power Supplies meet this safety requirement!)
Using the cooling option do not mix up the "in" and "out" plugs for the
cooling water.
Best cooling is obtained using a refrigerated circulator (chiller) with a
temperature of 5°C.
(Attention: Reduce flow rate to max. 0.5 - 1 l/min and use no organic
solvents or alcohol !)
Do not use alcohol (e.g. methanol, ethanol) or organic solvents for
cooling or cleaning the apparatus.
might
Fill the buffer tank with buffer up to the “--- max. fill line ---”. Use of
lower buffer volume can generate higher buffer temperature and
damage the instrument.
This products are designed and certified to meet EN 61010-1 safety
standards.
Certified products are safe to use when operated in accordance with
the instruction manual.
This instruments should not be modified or altered in any way.
Alteration of this instruments will void the warranty, void the
EN61010-1 certification, and create a potential safety hazard.
Eco Line Manual, Vers. 2014_06
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Operating Conditions:
max. V (DC)
max. mA
max. W
max. Temp.
Electrophoresis:
Eco-Mini:
Buffer tank E
Buffer tank EB
Buffer tank EBC*
200
200
200
Eco-Maxi:
Buffer tank EB
Buffer tank EBC*
400
400
50 (2 or 4 gels)
50 (2 or 4 gels)
50 (2 or 4 gels)
125 (2 gels)
250 (2 gels)
10
10
10
50°C
50°C
50°C
50
100
50°C
50°C
Max. power rating for electrophoresis without cooling should not be longer
than 18h at 5 W / gel for Eco-Mini and 18 h at 25 W / gel for Eco-Maxi.
Max. power rating for electrophoresis with cooling should not be longer than
18 h at 5 W / gel for Eco-Mini and 18 h at 50 W / gel for Eco-Maxi.
Blotting:
Eco-Mini:
Buffer tank EB
Buffer tank EBC*
100
100
300
300
30
30
50°C
50°C
Eco-Maxi:
Buffer tank EB
Buffer tank EBC*
60
100
1,000
1,000
60
130
50°C
50°C
Transfer time for blotting without cooling should not be longer than 4 h at 30 W
for Eco-Mini and 6 h at 60 W for Eco-Maxi.
Transfer time for blotting with cooling should not be longer than 18 h at 30 W
for Eco-Mini and 18 h at 130 W for Eco-Maxi.
* using the cooling option
Eco_Line Manual, 2014_06
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3
3.1
Setting-Up Operation
Scope of Delivery
Eco-Line is available in multiple versions for electrophoresis and/or blotting:
Eco-Mini
Complete system with
- buffer tank E, EB or EBC,
- Bigfoot Safety Lid with attached power cords,
- Electrophoresis Module,
- 2 sets of glass plates with fixed 1 mm spacers,
- 1 Dummy plate,
- 2 combs (10 wells) and
- manual
Eco-Mini System comes additionally with a Casting Stand for up to 2 single-gel or doublegel sandwiches.
Tankblot Eco-Mini
Complete modular system with
- buffer tank EB or EBC (with cooling base),
- Bigfoot Safety Lid with attached power cords,
- Blot Module,
- 4 Blotting Cassettes (colour-coded),
- 8 foam pads and
- manual
Eco-Maxi
Complete system with
- buffer tank EB or EBC,
- Bigfoot Safety Lid with attached power cords,
- Electrophoresis Module,
- 2 sets of glass plates with fixed 1 mm spacers,
- 2 combs (12 wells) and
- manual
Eco-Maxi System comes additionally with a Casting Stand for up to 2 gels.
Tankblot Eco-Maxi
Complete modular system with
- buffer tank EB or EBC (with cooling base),
- Bigfoot Safety Lid with attached power cords,
- Blot Module,
- 2 Blotting Cassettes (colour-coded),
- 4 foam pads and
- manual
Eco Line Manual, Vers. 2014_06
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Eco_Line Manual, 2014_06
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Spacer plate
Divider-Plate
Notched plate
Double-gel sandwich for Eco-Mini
Eco Line Manual, Vers. 2014_06
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Eco_Line Manual, 2014_06
15
3.2
Unpack and Check
Unpack and carefully examine the Eco-Line apparatus. Examine for signs of damage and
check the contents against the packing list. Report any damage to BIOMETRA. Do not
attempt to operate this device if physical damage is present.
Please keep the original packing material for return shipment in case of service issues
!! Attention !!
Please fill out and send back the warranty registration card. This is important for you
to claim full warranty.
3.3
Installation Conditions
Place the chamber in proximity to the Power Pack with which it is to be connected. Be sure
to place the chamber in a safe, dry location away from the edge of the working surface.
3.4
Connecting Conditions
The Eco-Mini Family apparatus have been designed to operate with D.C. current.
Warning: The apparatus must not be earthed.
Eco Line Manual, Vers. 2014_06
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4
Components
4.1
Systems Overview
Eco-Mini System E, EB or EBC
Eco-Maxi System EB or EBC

Buffer Tank E, EB or EBC
(Picture: Buffer Tanks Eco-Mini)

Bigfoot Safety Lid with power cords
(Picture: Bigfoot Safety Lid Eco-Mini)

Electrophoresis Module
(Picture: Electrophoresis Module Eco-Mini (left) and Eco-Maxi (right))

Casting stand
Picture: (Casting Stand Eco-Mini (left) and Eco-Maxi (right))
Eco_Line Manual, 2014_06
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Tankblot Eco-Mini or Eco-Mini C
Tankblot Eco-Maxi or Eco-Maxi C

Buffer Tank EB or EBC
(Picture: Buffer Tanks Eco-Mini)

Bigfoot safety lid with power cords
(Picture: Bigfoot Safety Lid Eco-Mini)

Blot Module
(Picture: Blot Module Eco-Mini (left)and Eco-Maxi (right))

Blotting Cassettes (colour coded)
(Picture: Blotting Cassette Eco-Mini (left) and Eco-Maxi (right))
Eco Line Manual, Vers. 2014_06
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4.2
Compatibility Overview
Buffer tank E for electrophoresis (Eco-Mini only)
Buffer tank EB for electrophoresis and blotting
Buffer tank EBC for electrophoresis and blotting;
with integrated cooling
Eco_Line Manual, 2014_06
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5
Electrophoresis
5.1
Gel Cassette Assembly
Clean and dry glass plates before use!


Single-gel sandwiches: Select a spacer plate of the desired gel thickness and place
the notched glass plate on top of it.
Double-gel sandwiches: Select a spacer plate and divider-Plate of the desired gel
thickness. Place the Devider-Plate on top of the spacer plate and the notched glass
plate on top of the Divider-Plate.
Spacer plate
Divider-Plate
Notched plate


Prepare a second assembly if two gels will be needed.
Slide the two glass plates assemblies (single-gel or
double-gel sandwiches) or one assembly and one
dummy plate into the Electrophoresis Module, keeping
the notched glass plate faced to the front of the module
and the spacer plate faced to the wall of the buffer tank:

Ensure that the glass plates of each assembly are flush on a level surface (not in
the Casting Stand!).
Leaking may occur if the plates are misaligned or damaged !

Tighten side clamps of the Electrophoresis Module with the white screws before the
Electrophoresis Module with the glass plate assemblies is placed into the Casting
Stand:
Please take care that the screws are tightened alternating and strong enough.
In case that the clamps are too loose the gel assembly could get out of place
during gel casting.
Eco Line Manual, Vers. 2014_06
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For Eco-Mini there is only 1 screw per clamp.
For Eco-Maxi there are 3 srews per clamp. Make sure that the screws in the middle
of each clamp are tightened first. After this tighten the diametrically opposed
screws crosswise.


Check that all glass plates are flush at the bottom.
Tighten the Dummy plate in a similar way at the opposite side of the Electrophoresis
Module in case of casting only a single gel.
The black Casting Stand gaskets must be clean and dry! Do not soak the
gaskets for prolonged periods prior to casting gels.
Take care that the black gaskets are exactly placed within the cavities in the stand and that
they are inserted evenly.
Pull out both eccentric screws of the Casting Stand.
Eco_Line Manual, 2014_06
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
Place the Electrophoresis Module with the glass plates assemblies straight down into
the Casting Stand.

Push the knob on each side of the Casting Stand
with the flat bottom of the eccentric interlocking pin
downwards.

Now simultaneously turn the eccentric screws on
both sides of the Casting Stand for approx. half a
turn.
Note: Don´t turn the eccentric screw more than
approx. 180°! Otherwise the glass plates can be
shifted upwards and would get leaky.
5.2









Gel Casting
Place a comb into the assembled gel cassette (one comb per single-gel sandwich, 2
combs per double-gel sandwich).
Mark the glass plate approx. 5 to 10 mm below the comb teeth.
Remove the comb(s) and pour the running gel (resolving gel) up to this level.
Prepare the running gel monomer solution by adding all reagents except APS and
TEMED. Degas the solution under vacuum. (Do not use a sink water aspirator!)
Add APS and TEMED to the degassed solution directly before pouring the gel.
Use a glass or disposable plastic pipette and smoothly pour the solution up to the
mark to prevent it from mixing with air.
Overlay the solution with bidistilled water or water saturated n-butanol. Prevent
mixing.
Allow the gel to polymerise for approx. 30 to 60 minutes. Polymerisation is visible by
the formation of a sharp dividing line between gel and overlay.
Remove the overlay solution from the surface of the polymerised gel. (If using water
saturated n-butanol rinse the gel surface with bidistilled water. Do not leave the
Eco Line Manual, Vers. 2014_06
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






5.3





alcohol overlay on the gel for more than 1 h because this might dehydrate the top of
the gel.)
Prepare the stacking gel monomer solution by adding all reagents except APS and
TEMED. Degas the solution under vacuum. (Do not use a sink water aspirator!)
Dry the top of the running gel with a sheet a filter paper before pouring the stacking
gel.
Add APS and TEMED to the degassed solution directly before pouring the gel.
Use a glass or disposable plastic pipette and smoothly pour the solution until the top
of the notched glass plate is reached.
Insert the desired comb between the two glass plates.
Allow the gel to polymerise for approx. 30 to 60 minutes.
Gentle remove the comb after polymerisation and rinse the wells carefully with
bidistilled water or running buffer.
System Assembly and Sample Loading
After polymerisation, lift the Electrophoresis Module with the gel assemblies from the
Casting stand and slide into the selected buffer tank.
Fill the buffer tank with the running buffer to the indicated max. level line.
Fill the chamber formed by the assemblies and the Electrophoresis Module with
running buffer to just under the edge of the outer (spacer) glass plate (approx. 5 mm
from the top of the gel assembly). Make sure the buffer level is above the gap formed
by the “ears” of the notched glass plate.
Dissolve the samples in sample buffer and carefully load into the wells of the stacking
gel.
Set the bigfoot safety lid onto the unit. Because of its asymmetric design this will only
work in the proper orientation, connecting the electrode of the buffer tank (anode)
with the red power cord and the Electrophoresis Module (cathode) with the black
power cord.
Eco_Line Manual, 2014_06
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5.4



Electrophoresis
Connect the safety plugs to a suitable Power Supply (e.g. Standard Power Pack P25
or P25T).
Be aware that the plus pole (red) is connected to the lower buffer reservoir.
Apply power to the apparatus and begin electrophoresis:
Eco-Mini:
const. 10 mA per gel is recommended for the pre-run and const. 25 mA per
Gel is recommended for the separation run. The max. Voltage has to be
limited at 200 V. Run time is depending on buffer and gel composition and
should be approx. 60 min. to 120 min. for SDS-PAGE.
Please notice: Compared to single-gel sandwiches the run time for doublegel sandwiches is approx. 25% longer!
Eco-Maxi:
const. 15- 25 mA per gel is recommended for the pre-run and const. 50 - 60
mA per Gel is recommended for the separation run. The max. Voltage has to
be limited at 400 V. Run time is depending on buffer and gel composition and
should be approx. 4 – 6 hours for SDS-PAGE.
5.5




System Reassembly and Gel Removal
After electrophoresis is completed, turn off the Power Supply and disconnect the
safety plugs.
Remove the Bigfoot Safety Lid and slide out the Electrophoresis Module with the
assemblies by carefully bending the Module towards the side with the Eco-Line logo
before lifting.
Discard the running buffer.
Loosen side clamps and remove gel assemblies.
Eco Line Manual, Vers. 2014_06
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6
Tankblotting
6.1
Assembling the Gel Sandwich
For gel sandwich assembly, mounting of blotting cassettes inside the buffer
tank EB or EBC and connecting to a power supply make sure that the
membrane is placed at the side of the gel which is close to the anode
(positive electrode side, red-coloured) if the target molecule are negatively
charged.















Pre-cool transfer buffer over night at 4°C.
After electrophoresis, immediately remove the gel from the bottom plate. Tip the plate
up side down, starting at one edge allow the gel to roll off into the transfer buffer.
Pre-equilibrate the gel in cool transfer buffer for 15 minutes with gentle agitation to
remove SDS and salts. This prevents the gel to change size during the blotting
transfer and reduces heating effects.
Cut blotting paper (e.g. Whatman 3MM or GB005) to the size of the gel (or the size of
the cassette) and soak 2 x 3 layers of thin (0.34 mm) or 2 x 1 layer of thick (approx.
1mm) with transfer buffer.
Wearing gloves, cut the membrane to the size of the gel and blotting paper.
Mark the membrane to indicate the side that the samples will be on.
Wet the membrane according to the manufacturers’ recommendations, followed by
equilibration in transfer buffer for 15 minutes.
Soak 2 foam pads in transfer buffer.
Building up the gel sandwich is best done in a shallow plastic box:
Open the blotting cassette and place the cassette with the black side down on a flat
surface.
Place a foam pad pre-soaked with transfer buffer in the black half of the blotting
cassette, followed by 3 layers of thin or 1 layer of thick filter paper pre-soaked with
buffer.
Gently place the gel on top of the pre-soaked filter paper. Beginning at one end of
the gel, align the filter papers with the gel edge and slowly lower the other end,
driving out any bubbles. Use a clean glass rod wetted in transfer buffer to roll out any
trapped bubbles.
Carefully align and overlay the transfer membrane onto the gel in one smooth action.
Work from the centre and let the ends roll down. Never re-position the membrane as
some transfer may occur on contact. Use a clean glass rod wetted in transfer buffer
to roll out any trapped bubbles.
Add 3 layers of thin or 1 layer of thick filter paper pre-soaked with buffer on top of the
membrane.
Complete the sandwich with a foam pad pre-soaked in blotting buffer followed by the
red half of the blotting cassette.
Eco-Maxi
Eco_Line Manual, 2014_06
Eco-Mini
25


6.2







Insert the cassette into the Blotting Module. Take care that the red side of the blotting
cassette is located at the side of the Blotting Module with the red-coloured electrode
(Eco-Mini) or at the side of the Blot Module with the red-coloured screws for fixing the
electrode (Eco-Maxi).
Up to 2 cassettes can be inserted into the Blotting Module of Eco-Maxi and up to 4
cassettes can be inserted into the Blotting Module of Eco-Mini.
Setting up the Buffer Tank (EB or EBC)
Cool and degas the appropriate volume of transfer buffer:
approx. 2,100 ml for use with Eco-Mini or
approx. 6,100 ml for use with Eco-Maxi.
Fill the buffer tank half to two-third with transfer buffer.
Connect the cooling supply to the tank base connectors (max. 0.5 l/min., use hose
clips to secure the tubing) or place the system into a cold room.
Add a stir bar (approx. 5 cm in length) to the tank to maintain uniform temperature
and conductivity during the blotting transfer.
Adjust mixing device to max. 200 rpm.
Slide the Blotting Module into the buffer tank.
Insert up to 4 cassettes into the Blotting Module of Eco-Mini or
insert up to 2 cassettes into the Blotting Module and turn the locking handles on both
sides downwards at Eco-Maxi.



6.3
Add transfer buffer up to the marked level.
Place the Bigfoot Safety Lid on the buffer tank and connect the attached power cords
to an appropriate power supply.
Start the blotting transfer.
Transfer of more than one Gel
Transfer efficiencies might decrease if more than 1 blotting cassette is inserted. In this case,
transfer times have to be increased to get the same transfer efficiencies as if blotting a
single gel sandwich.
Eco Line Manual, Vers. 2014_06
26
6.4
Transfer Times and Conditions
The best transfer time has to be determined experimentally. There is no formula for
determining transfer tine since there are too many variables involved to give specific transfer
conditions.
To optimise transfer times it is useful to stain the gel as well as the membrane using a
general staining method (e.g. Coomassie blue or silver stain for protein gels and Ponceau
red for proteins on membranes) and to compare the results with an identical gel which has
not been blotted.
6.5




System Reassembly and Blot Removal
After electrophoresis is completed, turn off the Power Supply and disconnect the
safety plugs.
Remove the Bigfoot Safety Lid and slide out the Blotting Module with the blotting
cassettes by carefully bending the Module towards the side with the Eco-Line logo
before lifting.
Discard the running buffer.
Remove the blotting cassettes.
Eco_Line Manual, 2014_06
27
7
7.1
Buffers and Staining/Destaining Solutions
Gel Solutions:
Running Gel
Eco-Mini
(5.5 ml per 0.75 mm gel)
Percent Gel
30:0.8 Acrylamide/Bis (ml)
1.88 M Tris/HCl, pH 8.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (l)
10% APS (l)
7.5%
1.4
1.1
1.1
1.9
5
28
10.0%
1.8
1.1
1.1
1.5
5
28
12.5%
2.3
1.1
1.1
1.0
5
28
15.0%
2.8
1.1
1.1
0.5
5
28
17.5%
3.2
1.1
1.1
0.0
5
28
(7 ml per 1 mm gel)
Percent Gel
30:0.8 Acrylamide/Bis (ml)
1.88 M Tris/HCl, pH 8.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (l)
10% APS (l)
7.5%
1.7
1.4
1.4
2.5
6
35
10.0%
2.3
1.4
1.4
1.9
6
35
12.5%
2.9
1.4
1.4
1.3
6
35
15.0%
3.5
1.4
1.4
0.7
6
35
17.5%
4.1
1.4
1.4
0.1
6
35
(10.5 ml per 1.5 mm gel)
Percent Gel
30:0.8 Acrylamide/Bis (ml)
1.88 M Tris/HCl, pH 8.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (l)
10% APS (l)
7.5%
2.6
2.1
2.1
3.7
9
53
10.0%
3.5
2.1
2.1
2.8
9
53
12.5%
4.4
2.1
2.1
1.9
9
53
15.0%
5.3
2.1
2.1
1.1
9
53
17.5%
6.1
2.1
2.1
0.2
9
53
Eco Line Manual, Vers. 2014_06
28
Running Gel
Eco-Maxi
(25 ml per 0.75 mm gel)
Percent Gel
30:0.8 Acrylamide/Bis (ml)
1.88 M Tris/HCl, pH 8.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (l)
10% APS (l)
7.5%
6.1
5.0
5.0
8.9
21
125
10.0% 12.5% 15.0% 17.5%
8.2
10.4
12.5
14.6
5.0
5.0
5.0
5.0
5.0
5.0
5.0
5.0
6.8
4.6
2.5
0.4
21
21
21
21
125
125
125
125
(33 ml per 1 mm gel)
Percent Gel
30:0.8 Acrylamide/Bis (ml)
1.88 M Tris/HCl, pH 8.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (l)
10% APS (l)
7.5%
8.0
6.6
6.6
11.8
28
165
10.0% 12.5% 15.0% 17.5%
10.8
13.7
16.5
19.3
6.6
6.6
6.6
6.6
6.6
6.6
6.6
6.6
8.9
6.1
3.3
0.5
28
28
28
28
165
165
165
165
(49 ml per 1.5 mm gel)
Percent Gel
30:0.8 Acrylamide/Bis (ml)
1.88 M Tris/HCl, pH 8.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (l)
10% APS (l)
7.5%
11.9
9.8
9.8
17.5
42
245
10.0% 12.5% 15.0% 17.5%
16.1
20.3
24.5
28.7
9.8
9.8
9.8
9.8
9.8
9.8
9.8
9.8
13.3
9.1
4.9
0.7
42
42
42
42
245
245
245
245
Eco_Line Manual, 2014_06
29
Stacking Gel, 5%
Eco-Mini
Gel Thickness
Gel Volume
Acrylamide/Bis 30:0.8 (ml)
0.625 M Tris/HCl pH 6.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (µl)
10% APS (µl)
0.75 mm
1.5 ml
0.2
0.3
0.3
0.7
2
8
1.0 mm
2 ml
0.3
0.4
0.4
0.9
2
10
1.5 mm
2.5 ml
0.4
0.5
0.5
1.1
3
13
0.75 mm
2.5 ml
0.3
0.5
0.5
1.2
3
13
1.0 mm
3.5 ml
0.5
0.7
0.7
1.6
4
18
1.5 mm
5.0 ml
0.8
1.0
1.0
2.2
6
26
Eco-Maxi
Gel Thickness
Gel Volume
Acrylamide/Bis 30:0.8 (ml)
0.625 M Tris/HCl pH 6.8 (ml)
0.5% SDS (ml)
dist. H2O (ml)
TEMED (µl)
10% APS (µl)
Attention:
The ammonium persulphate solution (10%) should be prepared fresh every day and
should be stored at 4C (refrigerator).
Eco Line Manual, Vers. 2014_06
30
7.2
Electrophoresis Buffer solutions:
SDS-Sample Buffer
for SDS-PAGE gels under reducing conditions
2 ml
0.2 g
5 ml
0.5 ml
0.1 ml
2.4 ml
0.625 M Tris/HCl pH 6.8
SDS
Glycerol
ß-Mercaptoethanol
Bromophenol Blue (1% in ethanol)
dist. H2O
Attention:
Dilute the sample buffer 1 to 2 with the sample (= 1 vol. buffer + 1 vol. sample)
Running Buffer pH 8.3 (5 liter):
15.1 g
72.0 g
5.0 g
Tris-Base
Glycine
SDS
Running Buffer pH 8.3 (10 liter):
30.2 g
144.0 g
10.0 g
Tris-Base
Glycine
SDS
Eco_Line Manual, 2014_06
31
7.3
Blotting Buffer Solutions:
Most transfer buffers for SDS PAGE gels are Tris/Glycin buffers (pH=8.3). At this pH
the negatively charged proteins will move to the anode. Using a transfer buffer with
acidic pH will return the moving direction because the positively charged proteins will
move to the cathode.
Typical transfer buffers are:
 20 mM Tris / 150 mM Glycine (pH=8.3),
 7.5 mM Tris / 1.2 mM Boric Acid (pH=8.9) or
 25 mM Sodium Phosphate (pH=6.5)
Towbin-buffer is the most commonly used buffer. It works well for transfer over night
(12 h) at const. 100 mA.
Towbin Buffer:
20 – 25 mM
150 – 192 mM
15 - 25 %
0.02 – 0.1 %
pH = 8.1 – 8.5
Tris-Base
Glycin
Methanol
SDS
The pH of Towbin-buffer will not be adjusted and is dependent on the quality of the
water and chemicals. The buffer should be degassed prior to the addition of SDS
and cooled to 4°C.
Higher concentration of SDS will increase transfer efficiency of proteins from the gel.
However, this might lower binding capacity of the membrane.
A good buffer with higher ion strength for more quick transfer rates is 50 mM CAPS
at pH=9.2. The pH of this buffer has to be adjusted with NaOH !
A very good buffer for quick transfer is 25 mM Na2HPO4 at pH=9.0 (+/- 0.1). This
buffer should be used at const. 300 mA.
A special transfer buffer for acidic gels with basic proteins, IEF gels and urea gels is
0.7% Acetic Acid.
Using acetic acid, the transfer direction is reverse (from “+” to “–“) and
transfer times are approx. 50% shorter than with use of “standard”
transfer buffers like Towbin Buffer!
A special transfer buffer for Glycoproteins, Polysaccharides and Lipoproteins is
10 mM Sodium Borate pH=9.2.
Eco Line Manual, Vers. 2014_06
32
7.4
Staining and Destaining solutions:
Staining Solution (1 liter)
2.0 g
0.5 g
425 ml
50 ml
100 ml
425 ml
Coomassie Brilliant Blue R 250
Coomassie Brilliant Blue G 250
Ethanol
Methanol
Acetic Acid
bidest. Water
Attention:
Stir overnight; filtrate before use; store in dark bottle!
Destaining Solution
fast destaining in:
45% Ethanol
10% Acetic Acid
75% bidest. Water
slow
25%
10%
45%
destaining in:
Isopropanol
Glacial Acid
bidest. Water
final destaining in:
7%
Acetic Acid in bidest. water
Eco_Line Manual, 2014_06
33
8
8.1
Trouble Shooting
Electrophoresis
Problem
Cause
Corrective Actions
Ensure that the two glass
plates of the assembly are
flush on a level surface.
Glass plates or spacers are dirty. Clean thoroughly.
Uneven/damaged edges of glass Use only glass plates that are
plates.
not damaged.
Gasket of Casting Stand is dirty. Clean thoroughly with a mild
detergent and rinse with
distilled water. Dry without
smearing.
Gasket of Casting Stand is
Replace gasket
damaged.
Gaskets of the Electrophoresis
Remove gaskets, wash in
Electrophoresis Modul
Module are deformed, dirty or
warm water to remove excess
assembly is leaking
too old.
of salts, and place each
gasket back into its groove.
If the gaskets are to old or
brittle, they have to be
replaced.
Check buffer recipe and
Running time is too long Buffer is too much diluted.
dilution. Try again with fresh
buffer.
Electrophoresis Modul assembly See above.
is leaking.
Running at too low current or
Make sure you are running
voltage.
using the suggested running
conditions.
For running at const. current,
the current value is per gel.
Buffer is too concentrated.
Check buffer recipe and
Running too fast
dilution. Try again with fresh
buffer.
If voltage is lower than usual
(when running at const.
current) the buffer is probably
too concentrated.
Current of voltage set too high.
Turn down electrical settings.
Turn down electrical settings.
Smiling of the dye front Center of the gel is hotter than
the sides.
Leakage of gel solution
Glass plates have not been
aligned correctly
Eco Line Manual, Vers. 2014_06
34
Problem
Cause
Corrective Actions
Load samples as quick as
possible and start the
electrophoresis run a.s.a.p.
after loading.
Diffusion of sample during run in Increase % of stacking gel or
the stacking gel.
increase current up to 25%
for the run in the stacking gel.
Lower ionic strength of sample. Match the ionic strength of
the sample with that of the
gel.
De-salt the sample. Use
Bands are narrower than Ionic strength of the sample is
higher
than
ionic
strength
of
the
sample buffer of the same
sample wells
gel.
ionic strength as the gel.
Bands spreading
outwards
Diffusion of sample when
loading.
Broad lanes at bottom of Samples with different buffers
are loaded.
gel
(normal in gradient gels.)
Gel not polymerised properly at
Skewed bands
the wells.
Salt concentration in the sample
is too high.
Separation gel is uneven at the
top.
The Electrophoresis Module
assembly is leaking through the
gel and/or along the side
spacers.
Streaked bands
Sample overloading.
Sample precipitation.
“Frowning” of outside
lanes
Eco_Line Manual, 2014_06
Leakage of buffer along the
sides.
Use identical sample buffer
for all samples loaded.
De-gas gel solution before
casting and/or increase APS
and TEMED concentration.
Control expiring date of the
used solutions.
De-salt samples.
Overlay the gel carefully
during polymerisation with
bidistilled water or water
saturated n-butanol.
Make sure the Casting stand
is levelled.
Check that the gel is a solid
gel inside the glass plates
and check the setup of the
apparatus to make sure that
sealing with the gasket is
perfect.
Check that spacer and com
thickness are identical.
Use lower concentration for
next run.
Centrifuge sample before
adding sample buffer and/or
use a lower % acrylamide gel.
Ensure that the two glass
plates of the assembly are
flush on a level surface.
35
Problem
Cause
Corrective Actions
Double bands
Re-oxidation or insufficient
reduction of the sample.
If using a reducing reagent
make sure that the buffer is
not too old. Increase the
concentration of 2-mercaptoethanol or dithiothreitol (DTT)
in the sample.
Eco Line Manual, Vers. 2014_06
36
8.2
Tankblotting
Problem
Cause
Corrective Actions
No transfer
Wrong order of membrane and
gel
Reverse order of membrane
and gel: gel on top of
membrane
Exchange connection:
Red = anode (+)
Black = cathode (-)
Place transfer membranes on
cathode side (lid) and gel on
anode side (base)
Increase transfer time
Wrong connection to the Power
Supply
Acidic buffer system: transfer
towards cathode!
Incomplete transfer
Irregular transfer
Proteins are stained on
both sides of the
membrane
Power Supply turns off
Strong increase of
voltage during transfer
Eco_Line Manual, 2014_06
Transfer time too short (check
by gel staining)
Transfer time too long: "blow
through" (i.e. proteins are
stained on both sides of the
membrane)
Air bubbles in the blot sandwich
Generation of air bubbles with
long transfer times by
electrolysis and heating up
Irregular current flow as size of
blotting papers and transfer
membrane are larger than gel
size
Blotting paper partially dried
Insufficiently soaked transfer
membrane
Spoiled electrodes
Transfer time to long: "blow
through"
Reduce transfer time. Use
nitrocellulose with smaller
pore size or PVDF membrane
Remove air bubbles by gently
rolling e.g. a pasteur pipette
over the membrane
Reduce transfer time, chill
buffer before use.
Transfer membranes and
blotting papers must be cut
exactly to gel size
Reduce transfer time
Check procedure for
prewetting membrane
Clean electrodes
Reduce transfer time. Use
nitrocellulose with smaller
pore size or PVDF membrane
Power Supply does not work at
Use proper Power Supply
low voltages (inner resistance
which can operate at low
too high)
voltages (e.g. Standard
Power Pack P25)
Blot sandwich dries out
Reduce amperage (soak
blotting paper cuts with more
transfer buffer)
Generation of air bubbles on the Chill buffers before use /
electrodes by electrolysis
Reduce transfer time
Insufficiently soaked transfer
Check procedure for
membrane
prewetting membrane
37
Problem
Cause
Corrective Actions
Intensive heating up
Amperage too high
Reduce amperage / Reduce
transfer time
Chill buffers before use /
Reduce transfer time
Check transfer buffers / Use
buffers described in this
manual
Transfer time too long
Conductivity of buffer too high
Eco Line Manual, Vers. 2014_06
38
9
Product Information
Apparatus (Eco-Mini):
017-100
Eco-Mini System E
Complete system with buffer tank E, Bigfoot Safety Lid, Electrophoresis
Module, 2 sets of glass plates with fixed 1 mm spacers, 1 Dummy plate, 2
combs (10 wells), Casting Stand and manual
Note: buffer tank E is not compatible with Blot Module 018-105
017-101
Eco-Mini E, dto., but without Casting Stand
017-102
Eco-Mini System EB
Complete modular system with buffer tank EB (without cooling base), Bigfoot
Safety Lid, Electrophoresis Module, 2 sets of glass plates with fixed 1 mm
spacers, 1 Dummy plate, 2 combs (10 wells), Casting Stand and manual
Note: buffer tank EB can be used with Blot Module 018-105
017-103
Eco-Mini EB, dto., but without Casting Stand
017-104
Eco-Mini System EBC
Complete modular system with buffer tank EBC (with cooling base), Bigfoot
Safety Lid, Electrophoresis Module, 2 sets of glass plates with fixed 1 mm
spacers, 1 Dummy plate, 2 combs (10 wells), Casting Stand and manual
Note: buffer tank EB can be used with Blot Module 018-105
017-105
Eco-Mini EBC, dto., but without Casting Stand
018-100
Tankblot Eco-Mini C
Complete modular system with buffer tank EBC (with cooling base), Bigfoot
Safety Lid, Blot Module, 4 Blotting Cassettes (colour-coded), 8 foam pads and
manual
Note: buffer tank EBC can be used with Gel Module 017-175
018-101
Tankblot Eco-Mini
Complete modular system with buffer tank EB (without cooling base), bigfoot
lid, Blot Module, 4 colour-coded Blotting Cassettes, 8 foam pads and manual
Note: buffer tank EBC can be used with Gel Module 017-175
Eco_Line Manual, 2014_06
39
Accessories (Eco-Mini):
017-180
Casting Stand (two-place) for Eco-Mini
017-169
017-170
017-171
Buffer tank E for Eco-Mini; without lid; for electrophoresis only
Buffer tank EB for Eco-Mini; without lid; for electrophoresis and blotting
Buffer tank EBC for Eco-Mini; without lid; for electrophoresis and blotting,
with cooling base
017-172
Bigfoot Safety Lid for Eco Mini; fits to all buffer tanks
017-175
Electrophoresis Module for Eco-Mini; for 2 gels; fits to all buffer tanks
017-120
017-121
017-122
017-125
017-127
Glass plate for Eco-Mini; with fixed spacers, 1mm
Glass plate for Eco-Mini; with fixed spacers, 0.75mm
Glass plate for Eco-Mini; with fixed spacers, 1.5mm
Notched glass plate for Eco-Mini
Dummy plate for Eco-Mini
017-183
017-184
017-185
Divider-Plate with fixed spacers, 1.0 mm
Divider-Plate with fixed spacers, 0.75 mm
Divider-Plate with fixed spacers, 1.5 mm
017-187
017-188
017-189
Divider-Plate Set (2 divider-Plates and 2 combs, 10 well), 1.0 mm
Divider-Plate Set (2 divider-Plates and 2 combs, 10 well), 0.75 mm
Divider-Plate Set (2 divider-Plates and 2 combs, 10 well), 1.5 mm
017-130
017-131
017-132
017-133
017-134
Comb for Eco-Mini; 9 teeth, 0.75mm, MTP compatible
Comb for Eco-Mini; 10 teeth, 0.75mm
Comb for Eco-Mini; 12 teeth, 0.75mm
Comb for Eco-Mini; 15 teeth, 0.75mm
Preparative comb for Eco-Mini; 1 well plus 2 marker wells, 0.75mm
017-140
017-141
017-142
017-143
017-144
Comb for Eco-Mini; 9 teeth, 1.0mm, MTP compatible
Comb for Eco-Mini; 10 teeth, 1.0mm, MTP compatible
Comb for Eco-Mini; 12 teeth, 1.0mm, MTP compatible
Comb for Eco-Mini; 15 teeth, 1.0mm, MTP compatible
Preparative comb for Eco-Mini; 1 well plus 2 marker wells, 1.0mm
017-150
017-151
017-152
017-153
017-154
Comb for Eco-Mini; 9 teeth, 1.5mm, MTP compatible
Comb for Eco-Mini; 10 teeth, 1.5mm, MTP compatible
Comb for Eco-Mini; 12 teeth, 1.5mm, MTP compatible
Comb for Eco-Mini; 15 teeth, 1.5mm, MTP compatible
Preparative comb for Eco-Mini; 1 well plus 2 marker wells, 1.5mm
018-105
Blot Module for Tankblot Eco-Mini
018-111
Blotting cassette for Tankblot Eco-Mini; colour-coded, for 1 mini
gel
Foam pads for Tankblot Eco-Mini; 4 pcs (for two cassettes)
018-113
Eco Line Manual, Vers. 2014_06
40
Apparatus (Eco-Maxi):
017-400
Eco-Maxi System EB
Complete modular system with buffer tank EB (without cooling base), Bigfoot
Safety Lid, Electrophoresis Module, 2 glass plates with fixed 1.0 mm spacers,
2 notched glass plates, 2 combs (1 mm, 12 wells), Casting Stand and
manual
Note: buffer tank EB can be used with Blot Module 018-405
017-401
Eco-Maxi EB, dto., but without Casting Stand
017-402
Eco-Maxi System EBC
Complete modular system with buffer chamber EBC (with integrated cooling
base), Bigfoot Safety Lid, Electrophoresis Module, 2 glass plates with fixed
1.0 mm spacers, 2 notched glass plates, 2 combs (1 mm, 12 wells), Casting
Stand and manual
017-403
Eco-Maxi EBC, dto., but without Casting Stand
018-400
Tankblot Eco-Maxi C
Complete modular system with buffer chamber EBC (with integrated cooling
base), Bigfoot Safety Lid, 2 Blotting Cassettes (colourcoded, each for 1 gel
with size 22 cm x 19 cm), 4 foam pads and manual
Note: buffer tank EBC can be used with Electrophoresis Module 017-475
018-401
Tankblot Eco-Maxi
Complete modular system with buffer chamber EB (without cooling base),
Bigfoot Safety Lid, 2 Blotting Cassettes (colour-coded, each for 1 gel with size
22 cm x 19 cm), 4 foam pads and manual
Note: buffer tank EB can be used with Electrophoresis Module 017-475
Eco_Line Manual, 2014_06
41
Accessories (Eco-Maxi):
017-480
Casting Stand (two-place) for Eco-Maxi
017-471
017-472
Buffer tank EB for Eco-Maxi; without bigfoot safety lid
Buffer tank EBC for Eco-Maxi; without bigfoot safety lid
017-474
Bigfoot Safety Lid for Eco-Maxi; fits to all buffer tanks
017-475
Electrophoresis Module for Eco-Maxi; for 2 gels
017-420
017-421
017-423
017-425
017-426
Glass plate for Eco-Maxi; with fixed spacers, 1 mm
Glass plate for Eco-Maxi; with fixed spacers, 0.75 mm
Glass plate for Eco-Maxi; with fixed spacers, 1.5 mm
Notched glass plate for Eco-Maxi
Dummy plate for Eco-Maxi
017-430
017-431
017-432
017-433
017-434
Comb for Eco-Maxi; 12 teeth, 0.75 mm
Comb for Eco-Maxi; 19 teeth, 0.75 mm, MTP compatible
Comb for Eco-Maxi; 25 teeth, 0.75 mm
Comb for Eco-Maxi; 30 teeth, 0.75 mm
Preparative comb for Eco-Maxi; 1 well plus 2 marker wells, 0.75 mm
017-440
017-441
017-442
017-443
017-444
Comb for Eco-Maxi; 12 teeth, 1.0 mm
Comb for Eco-Maxi; 19 teeth, 1.0 mm, MTP compatible
Comb for Eco-Maxi; 25 teeth, 1.0 mm
Comb for Eco-Maxi; 30 teeth, 1.0 mm
Preparative comb for Eco-Maxi; 1 well plus 2 marker wells, 1.0 mm
017-450
017-451
017-452
017-453
017-454
Comb for Eco-Maxi; 12 teeth, 1.5 mm
Comb for Eco-Maxi; 19 teeth, 1.5 mm, MTP compatible
Comb for Eco-Maxi; 25 teeth, 1.5 mm
Comb for Eco-Maxi; 30 teeth, 1.5 mm
Preparative comb for Eco-Maxi; 1 well plus 2 marker wells, 1.5 mm
018-405
Blot Module for Tankblot Eco-Maxi, for up to 2 Blotting Cassettes
(each for 1 gel with size 22 cm x 19 cm) and 4 foam pads
018-411
Blotting cassette, colour coded (black/red) for Tankblot Eco-Maxi
Blot Module, 1 pcs
Foam pads for Tankblot Eco-Maxi; 4 pcs (for two cassettes)
018-413
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Blotting Paper:
10 426 981
10 426 994
Whatman GB005, 200 mm x 200 mm, 1.2 mm thick, 25 sheets
Whatman GB005, 580 mm x 580 mm, 1.2 mm thick, 25 sheets
3017915
3030 931
Whatman 17Chr, 460 mm x 570 mm, 0.92 mm thick, 100 sheets
Whatman 3MM, 580 mm x 680 mm cm, 0.34 mm thick, 100 sheets
Nitrocellulose Blotting Membranes:
10 401 396
Whatman Protran-BA83, Pore size 0.20 µm, 30 cm x 3 m roll
10 401 196
10 401 180
10 402 580
Whatman Protran-BA85, Pore size 0.45 µm, 30 cm x 3 m roll
Whatman Protran-BA85, Pore size 0.45 µm, 30 cm x 60 cm, 5 sheets/pack
Whatman Protran-BA85, Pore size 0.45 µm, 33 cm x 56 cm, 5 sheets/pack
10 439 196
Whatman Protran-BA-S 85, Pore size 0.45 µm, 30 cm x 3 m roll
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Eco Line Manual, Vers. 2014_06
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10
10.1
Maintenance and Repair
Cleaning and Maintenance
Rinse buffer tank, bigfoot lid, Electrophoresis Module, Blot Module, Blotting cassettes and
Casting Stand thoroughly with running water and finally with distilled water after every use.
It is recommended to allow the chamber and Electrophoresis or Blotting Module to air dry
rather than drying with a towel to avoid damage of the electrode.
Glass plates and combs should be washed with a laboratory detergent and then rinsed
thoroughly with distilled water.
Avoid extended submersion of spacer plates in water or cleaning solution. Limit submersion
to time used for cleaning. Prolonged submersion compromises the integrity of the adhesive.
Do not use alcohol >10% (e.g. methanol, ethanol) or organic solvents (e.g. acetone) for
cleaning of electrodes or housing as this will cause acrylic to crack.
The system should never be autoclaved or placed in a microwave.
10.2
Servicing
Regular servicing is not necessary. Nevertheless cleaning of the electrodes should be done
using a damp cloth wetted with dest. water after each transfer.
In case that the electrodes are clogged after longer time of use and the blotting transfer is
inhomogeneous or reduced, please contact the Biometra Service Department.
10.3
Replacement of Spare Parts
Only original spare parts mentioned in these operating instructions are allowed.
10.4
Other Accessories
See chapter 8 (Product Informations) for further details.
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11
Service
Should you have any problems with this unit, please contact our service department or your
local Biometra dealer:
Biometra GmbH
Service Department
Rudolf-Wissell-Straße 14 - 16
D-37079 Goettingen
Phone: +49 (0)5 51 50 68 6 - 10 or 12
Fax: +49 (0)5 51 50 68 6 -11
e-mail: [email protected]
If you would like to send the unit back to us, please read the following return
instructions.
11.1







Instructions for Return Shipment
Return only defective devices. For technical problems which are not definitively
recognisable as device faults please contact the Technical Service Department at
Biometra (Tel.: +49 (0)5 51-50 88 1-10 or -12, Fax: +49 (0)5 51-50 88 1-11, e-mail:
[email protected]).
Please contact our service department for providing a return authorization number
(RAN). This number has to be applied clearly visible to the outer box. Returns without
the RAN will be not be accepted!
Important: Carefully clean all parts of the instrument from residues, and of biologically
dangerous, chemical or radioactive contaminants. If an instrument is
contaminated, Biometra will be forced to refuse to accept the device. The
sender of the repair order will be held liable for possible damages and losses
resulting from insufficient decontamination of the device.
Please prepare written confirmation (use the “Equipment Decontamination
Declaration” following on the next page) that the device is free of biologically dangerous,
chemical or radioactive contaminants. This confirmation must be attached to the outside
of the packaging.
Use the original packing or a similarly robust packing when returning the device. If not
available, contact Biometra or your local distributor.
Label the outside of the box with “CAUTION! SENSITIVE INSTRUMENT!” and the RAN
number sticker. Attach the Decontamination Declaration!
Please enclose a note which contains the following:
a) Sender’s name and address,
b) Name of a contact person for further inquiries with telephone number.
c) Precise description of the fault, which also reveals during which procedures the
fault occurred, if possible.
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12
Equipment Decontamination Certificate
To enable us to comply with German law (i.e. §71 StrlSchV, §17 GefStoffV and §19 ChemG)
and to avoid exposure to hazardous materials during handling or repair, please complete this
form, prior to the equipment leaving your laboratory.
COMPANY / INSTITUTE
__________________________________________________
ADDRESS
______________________________________________________________
PHONE NO
_________________________
E-MAIL
______________________________________________________________
EQUIPMENT
FAX NO_________________________
Model
______________
Serial No
______________
______________
______________
______________
______________
______________
______________
If on loan / evaluation Start Date: __________________ Finish Date __________________
Hazardous materials used with this equipment:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
Method of cleaning / decontamination:
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
The equipment has been cleaned and decontaminated:
NAME __________________________________ POSITION _______________________
(HEAD OF DIV./ DEP./ INSTITUTE / COMPANY)
SIGNED ________________________________ DATE ____________________________
PLEASE RETURN THIS FORM TO BIOMETRA GMBH OR YOUR LOCAL BIOMETRA
DISTRIBUTOR TOGETHER WITH THE EQUIPMENT.
PLEASE ATTACH THIS CERTIFICATE OUTSIDE THE PACKAGING. INSTRUMENTS
WITHOUT THIS CERTIFICATE ATTACHED WILL BE RETURNED TO SENDER.
General Information for Decontamination:
Eco_Line Manual, 2014_06
47
Please contact your responsible health & safety officer for details.
Use of radioactive substances:
Please contact your responsible person for details.
Use of genetically change organism or parts of those:
Please contact your responsible person for details.
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13
Note for Disposal of Electric/Electronic Waste
This symbol (the crossed-out wheelie bin) means, that this product should be brought
to the return systems and/or separate systems available to end-users according to
yours country regulations, when this product has reached the end of its lifetime!
For details, please contact your local distributor!
This symbol applies only to the countries within the EEA*.
*EEA = European Economics Area, comprising all EU-members plus Norway, Iceland and
Liechtenstein.
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14
EC – Declaration of Conformity /
EU - Konformitätserklärung
Goettingen, January 2010
im Sinne der EG-Richtlinie für die elektromagnetische Verträglichkeit 2004/108/EG
following the EC directive about the electromagnetic compatibility 2004/108/EC
Hiermit erklären wir, dass folgende Elektrophorese und Blotting Systeme
Herewith we declare that the following Electrophoresis and Blotting Systems
Typ / type:
Eco-Mini E, Eco-Mini System E,
Eco-Mini EB, Eco-Mini System EB,
Eco-Mini EBC, Eco-Mini System EBC,
Eco-Maxi EB, Eco-Maxi System EB,
Eco-Maxi EBC, Eco-Maxi System EBC,
Tankblot Eco-Mini , Tankblot Eco-Mini C
Tankblot Eco-Maxi, Tankblot Eco-Maxi C
Best.-Nr. / Order No.
017-100, 017-101, 017-102, 017-103, 017-104, 017-105,
017-400, 017-401, 017-402, 017-403,
018-100, 018-101,
018-400, 018-401
den grundlegenden Anforderungen der
corresponds to the basic requirements of
EG-Niederspannungsrichtlinie 2006/95/EG entsprechen.
EC low voltage directive 2006/95/EC.
Juergen Otte PhD
Quality Manager
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15
Warranty
This laboratory instrument is produced with the highest practical standards of materials,
workmanship, and design. The design and manufacture of parts have been conceived with
one purpose - to produce units which will give satisfactory service.
Biometra GmbH guarantees this unit to be free from defects in materials or workmanship
under normal use or service for 24 month from date of shipment.
If, during this time, this unit proves defective in materials or workmanship, Biometra GmbH
will repair or replace it free of charge if returned to us prepaid.
This guarantee does not cover damage in transit, damage caused by carelessness, misuse
or neglect, or unsatisfactory performance as a result of conditions beyond our control; or
consequential losses as a result of failure of our product.
Biometra GmbH
Rudolf-Wissell-Str. 30
D-37079 Goettingen
Tel:
+49 (0)5 51 50 68 6-0
Fax:
+49 (0)5 51 50 68 6-66
e-mail: [email protected]
internet: http://www.biometra.de
Eco_Line Manual, 2014_06
Service Department
Rudolf-Wissell-Str. 14-16
D-37079 Goettingen
Tel.: +49 (0)5 51 50 68 6-10 or -12
Fax.: +49 (0)5 51 50 68 8-11
e-mail: [email protected]
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