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Eppendorf Eporator®
Operating manual
Copyright© 2013 Eppendorf AG, Hamburg. No part of this publication may be reproduced
without the prior permission of the copyright owner.
Eppendorf
Hamburg, Germany.
Eppendorf Eporator
Excel
States and other countries.
Registered trademarks are not marked in all cases with ™ or ® in this manual.
®
and the Eppendorf logo are registered trademarks of Eppendorf AG,
®
is a registered trademark of Eppendorf AG, Hamburg, Germany.
®
and Microsoft® are registered trademarks of Microsoft Corporation in the United
4309 900.010-04/082013
Table of contents
Eppendorf Eporator®
English (EN)
Table of contents
1 Operating instructions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.1 Hazard symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2.2 Degrees of danger . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.3 Symbols used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.4 Glossary. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2 Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
2.1 Main illustration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
2.2 Delivery package . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
2.3 Features. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.3.1 The principle of electroporation . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3 Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.1 Intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.2 User profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.3 Information on product liability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.4 Safety instructions on the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.5 Warnings for intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
4.1 Preparing installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
4.2 Selecting the location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
4.3 Installing the instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3
5 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5.1 Overview of operating controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
5.2 Recommendations for sample preparation . . . . . . . . . . . . . . . . . . . . . . . . . . 14
5.2.1 DNA preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
5.2.2 Electroporation medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
5.2.3 Growth and preparation of cells . . . . . . . . . . . . . . . . . . . . . . . . . . 15
5.2.4 Temperature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
5.3 Performing electroporation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
5.3.1 Switching on the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
5.3.2 Inserting the cuvette . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
5.3.3 Electroporation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
5.4 Regeneration of the cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
5.5 Determination of the transformation efficiency . . . . . . . . . . . . . . . . . . . . . . 18
5.6 Programs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
5.6.1 Loading program . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
5.6.2 Saving program . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
5.7 Enhanced settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
5.8 Exporting data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Table o f co nte nts
Eppendorf Eporator®
4
English (EN)
6 Troubleshooting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
6.1 General errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
6.2 Error messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
6.2.1 Operational error . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
6.2.2 Device error . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
7 Maintenance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
7.1 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
7.2 Replacing fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
8 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
8.1 Power supply. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
8.2 Ambient conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
8.3 Weight/dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
8.4 Interfaces. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
8.5 Operating mode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
9 Ordering Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
9.1 Eporator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
9.2 Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
10 Transport, storage and disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
10.1 Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
10.2 Decontamination before shipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
10.3 Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
10.4 Disposal. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Operating instructions
Eppendorf Eporator®
English (EN)
1 Operating instructions
1.1 Using this manual
Please read this operating manual completely before using the device for the first time.
Please view this operating manual as part of the product and keep it somewhere easily
accessible.
When passing the device on to third parties, be sure to include this operating manual.
If this manual is lost, please request another one. The current version can be found on
our website www.eppendorf.com
1.2 Danger symbols and danger levels
1.2.1 Hazard symbols
Toxic substances Electric shock
Hazard point Material damage
1.2.2 Degrees of danger
The degree of danger is a part of a safety note and distinguishes the possible results of
non-observance from each other.
DANGER Will lead to severe injuries or death.
WAR NIN G May lead to severe injuries or death.
CAUTION May lead to light to moderate injuries.
NOTICE May lead to material damage.
.
5
1.3 Symbols used
Representation Meaning
Handling
1.
2.
• List:
Text Name of fields in the software
Actions in the specified order
Useful information
Operating instructions
Eppendorf Eporator®
6
English (EN)
1.4 Glossary
A
Arcing
If electrical voltage is applied between two parallel electrodes, a current flows in an
evenly distributed layer. If the voltage exceeds a critical value, this layer contracts to a
narrow circuit with high current density: an electric arc. The electrode material melts at
this location, and explosive evaporation occurs. The cuvette can be destroyed under these
conditions.
E
Electrical field strength
Ratio of potential difference between two electrodes (in V) and the distance between
these electrodes (electrode gap; in cm). However, this only applies if the electrical field is
homogeneous, as with parallel plate electrodes (e.g., in Eppendorf Electroporation
Cuvettes).
T
Time constant
Time during which the voltage decreases to the value U/e.
2 Product description
1 2
3 4
56
2.1 Main illustration
Abb. 2-1:Main Illustration
Product description
Eppendorf Eporator®
English (EN)
7
Fig. 2-1: Main Illustration
1 Operating controls
2 Cuvette carrier
In the cuvette shaft
3 RS232 port
Only for Technical Service
2.2 Delivery package
Quantity Description
1 Eppendorf Eporator
1Mains/power cord
1 Cuvette holder
1 Eporator operating manual
4USB port
For inserting a USB stick
5Mains switch
6 Mains connection socket
Product description
Eppendorf Eporator®
8
English (EN)
2.3 Features
The Eporator is inserted for the electroporation. It contains a capacitor that is discharged
during electroporation using a resistor, thus generating an exponential discharge curve. A
voltage between 200 V and 2 500 V can be set. The exponential pulse generated by the
Eporator is transferred to a disposable electroporation cuvette that contains the biological
sample.
Unlike devices from other manufacturers, the Eporator features an integrated cuvette
carrier with cuvette holder.
The construction of the Eporator minimizes the risk of short circuits. This also applies in
the case of prohibited, high salt concentrations or maximum voltage. Even in the most
unlikely situation of an electric arc in the cuvette, no bacterial suspension can escape
from the cuvette and contaminate the device.
The Eporator is easy to operate. None of the device components require
user-maintenance.
The experimental electroporation data can be saved on a USB stick and evaluated on a
PC.
Application protocols for the electroporation of various bacteria and yeast strains can be
found on the Eppendorf home page www.eppendorf.com
.
2.3.1 The principle of electroporation
DNA and bacteria Electrical charge
200 V - 2 500 V;
approx. 5 ms
With the electroporation method, macromolecules such as DNA can be placed in
electrocompetent bacteria or yeast strains. In the process, small-volume samples with
high resistance are exposed to pulses with very high electrical field strengths. The short,
high voltage pulses create temporary holes or pores in the cell membrane, through which
macromolecules, e.g. plasmid DNA, can diffuse into the cell. The holes close after removal
of the electrical field and a period of regeneration. The inserted plasmid DNA can then be
transcribed and replicated in the cell.
Unlike chemical transformation, electroporation is characterized by high transformation
efficiency and simple execution.
DNA in bacteria
Eppendorf Eporator®
Safety
English (EN)
3Safety
3.1 Intended use
The Eporator is intended for indoor use only and enables the simple and safe
electroporation of bacteria and yeast strains using standard protocols.
3.2 User profile
This device must only be used by skilled personnel with the appropriate training.
Before using the device, read the operating manual carefully and familiarize yourself with
the device's mode of operation.
3.3 Information on product liability
In the following cases, the protection provided by the device may be impaired. The
liability for the function of the device passes to the operator if:
• The device is not used in accordance with the operating manual.
• The device is used outside of the range of application described in the preceding
chapters.
• The owner has made unauthorized modifications to the device.
3.4 Safety instructions on the device
Depiction Meaning
WAR NING
Follow the operating manual.
9
3.5 Warnings for intended use
WARNING! Damage to health from toxic, radioactive or aggressive chemicals
as well as infectious liquids and pathogenic germs.
Observe the national regulations for handling these substances, the biological
security level of your laboratory, the material safety data sheets and the
manufacturer's application notes.
Wear personal protective equipment (PPE).
Safety
Eppendorf Eporator®
10
English (EN)
WARNING! Lethal voltages inside the device.
Ensure that the housing is always closed and undamaged so that no parts
Do not remove the housing of the device.
Do not allow any liquids to enter the inside of the housing.
Do not allow the device to be opened by anyone except service personnel who
WARNING! Risk of electrical shock from damage to the device or mains
cable.
The device may only be switched on if the device and mains cable are
Only use devices which have been professionally installed or repaired.
WARNING! Danger from using an incorrect power supply.
Only connect the device to voltage supplies which correspond with the
Only use sockets with a protective grounding conductor and a suitable mains
inside the device can be contacted by accident.
have been specifically authorized by Eppendorf.
undamaged.
electrical requirements on the name plate.
cable.
CAUTION! Poor safety due to incorrect accessories and spare parts.
The use of accessories and spare parts other than those recommended by
Eppendorf may impair the safety, functioning and precision of the device.
Eppendorf cannot be held liable for any damage resulting from the use of
non-recommended accessories and spare parts or from the improper use of such
equipment.
Only use accessories and original spare parts recommended by Eppendorf.
NOTICE! Damage to device due to penetration of liquids.
Liquid can enter the device during electroporation with cuvettes without lids.
Only cuvettes with square lids may be used for electroporation.
Eppendorf Eporator®
Installation
English (EN)
4 Installation
4.1 Preparing installation
Retain the transport carton and packing material for subsequent safe transport
or storage.
Check all parts for any transport damage.
4.2 Selecting the location
Please select the location for the Eporator according to the following criteria:
• Mains connection (230 V/120 V/100 V) according to the name plate. This is located on
the rear side of the device.
• At least 10 cm away from adjacent devices and walls.
• Solid bench with stable, horizontal and even work surface.
4.3 Installing the instrument
WARNING! Danger from using an incorrect power supply.
Only connect the device to voltage supplies which correspond with the
electrical requirements on the name plate.
Only use sockets with a protective grounding conductor and a suitable mains
cable.
11
1. Connect the provided mains cable to the mains connection socket of the Eporator and
the power supply.
2. Switch on the Eporator at the mains power switch.
Installation
Eppendorf Eporator®
12
English (EN)
Eppendorf Eporator®
Start
exit
menu
enter
P2
P1
1 2
3
4
5
6
Operation
English (EN)
5Operation
5.1 Overview of operating controls
Familiarize yourself with the operating control elements and the display of the Eporator
before using it for the first time.
Abb. 5-1:Operating controls
Fig. 5-1: Operating controls
13
1 P1 program key with control LED
Press: load voltage value. Press and hold
(> 2 s): save current voltage value.
2 P2 program key with control LED
Press: load voltage value. Press and hold
(> 2 s): save current voltage value.
3 Press the Start key
Start electroporation
4Arrow keys
5Exit key
6 menu/enter key
Set the voltage
Exit the menu
Select the menu parameter
Operation
1
2
345
Eppendorf Eporator®
14
English (EN)
Abb. 5-2:Display
Fig. 5-2: Display
1 Actual voltage value (in V)
2 Actual discharge time (in ms)
3 Voltage symbol
The voltage symbol appears after
4 Cuvette symbol
The cuvette symbol appears when a
cuvette is inserted.
5 Set voltage (in V)
electroporation and disappears after the
cuvette carrier has been removed.
5.2 Recommendations for sample preparation
Independent of the device, the success of an electroporation is influenced by a variety of
factors:
• Quality and concentration of the inserted DNA
• Quality and concentration of the cells
• Resuspension medium of the DNA and cells
5.2.1 DNA preparation
• DNA quality: In order to obtain a high transformation efficiency, the DNA solution
should be pure and free of salts.
• Buffer: DNA dissolved in TE buffer is acceptable if the DNA was dissolved in approx.
ten times the quantity of electrocompetent cells.
• Salt concentration: DNA from enzyme reactions (e.g. ligation) can be immediately
used for electroporation if the salt concentration is under 5 M. If the ionic strength of
the reaction combination is too high, it can be reduced via dilution or ethanol
precipitation. After ethanol precipitation, the DNA can be resuspended in sterile,
demineralized water or TE buffer.
• Incubation: Before electroporation, do not incubate the DNA with the cell suspension
too long. Generally, the DNA should be added to the cells one minute before
electroporation and the solution should be incubated at 0 °C. Long incubation times
can lead to DNA degradation due to the DNases in the cell suspension.
Eppendorf Eporator®
Operation
English (EN)
• DNA concentration:The concentration of DNA can significantly influence the
transformation efficiency.
• Frequency and efficiency: The frequency is defined as the number of transformed
cells per surviving cells. The efficiency is defined as transformed cells per μg DNA. You
can obtain a high frequency by using high DNA concentrations. High efficiency is
achieved by using high cell concentration. Reducing the DNA concentration helps
prevent co-transformations of the same cell.
5.2.2 Electroporation medium
• Sensitivity of the cells: The cells are sensitive to external influences because
electroporation creates temporary pores in the cell membrane.
• Electrolysis of the medium: During electroporation, the electrolysis of the medium
significantly influences the characteristics of the medium (e.g. the pH value). Many
cells can die if fresh medium is not immediately added after electroporation for the
recovery of the cells (see Regeneration of the cells on p. 18).
• Ionic strength of the medium: The ionic strength of the medium must be taken into
account during electroporation of cells. In order to keep the resistance of the medium
as high as possible, salts must be removed from the cell and DNA preparation. Ions
remaining in the cell suspension often come from the culture medium. Higher
transformation efficiency can be obtained by removing salts from the DNA solution and
cell preparations. Generally, the lowest possible ionic solution that cells can withstand
is preferred.
15
5.2.3 Growth and preparation of cells
• Growth phase of cells: For optimum electroporation efficiency use bacterial (e.g. E.
coli) or yeast cells in the exponential growth phase.
• Preparation of cells: Thoroughly wash the cells in order to remove the growth medium
that affects electrocompetence.
• Concentration of cells: Use a final concentration of cells of about 1-3 x 10
11
cells/mL.
If this value is exceeded, the homogenity of the electrical field can be affected.
• Requirements for an electroporation: Each bacteria strain and yeast strain has
optimal conditions that must be determined empirically. These conditions include:
– The cell volume
– The quantity of the specific plasmid
– The used field strength (E). For E. coli, a field strength of 12-19 kV/cm is generally
required to reach a maximum transformation efficiency. The field strengths are
calculated from the voltage used and the distance of the electrodes (E = V/cm).
Operation
Eppendorf Eporator®
16
English (EN)
5.2.4 Temperature
• Cooling electroporation cuvettes: Electroporation of microorganisms produces the
best results at low temperatures (0-4 °C). Cool the electroporation cuvettes to 0 °C
before electroporation. Remove residual moisture from the electroporation cuvette
before inserting it in the Eporator.
5.3 Performing electroporation
5.3.1 Switching on the device
Press the mains power switch on the rear of the device to switch on the device.
5.3.2 Inserting the cuvette
To increase the efficiency of the electroporation, the electroporation cuvette can
be cooled prior to filling. Remove residual liquids from the cuvette before further
use.
The integrated cuvette holder is located at the front of the device to the right of the control
panel.
Proceed as follows:
1. Remove the cuvette holder from the device.
The cuvette holder is now freely accessible for the insertion of the electroporation
cuvette.
2. Remove the electroporation cuvette from the individual packaging.
3. Remove the lid of the electroporation cuvette.
4. Fill the electroporation cuvette with the sample. The slit between the plate electrodes
must be filled without bubbles.
5. Place the locking lid on the electroporation cuvette.
NOTICE! Damage to device due to penetration of liquids.
Liquid can enter the device during electroporation with cuvettes without lids.
Only cuvettes with square lids may be used for electroporation.
6. Insert the electroporation cuvette into the cuvette holder with the plastic nose pointing
towards the back.(Fig. 5-3 on p. 17)
Eppendorf Eporator®
Operation
English (EN)
Abb. 5-3:Insert the electroporation cuvette
Fig. 5-3: Insert the electroporation cuvette
Operate the cuvette holder with two hands to prevent the cuvette holder from
tipping.
7. Slide the holder in the cuvette shaft of the Eporator until it engages (Fig. 5-4 on p. 17).
Abb. 5-4:Insert the cuvette holder
17
Fig. 5-4: Insert the cuvette holder
In the display, the actual parameters of the last run disappear and a cuvette symbol
appears in the lower line.
Operation
Start
Eppendorf Eporator®
18
English (EN)
5.3.3 Electroporation
1. Set a voltage between 200 V and 2 500 V using the arrow keys.
After switching on the device, the last set voltage is always displayed. The
most frequently used voltages can be saved and accessed using the program
keys (see Programs on p. 19).
2. Press the Start key to start the electroporation process.
• Charge and a progress bar appear in the display during loading.
• A signal tone sounds after the discharge.
• After the electroporation, the actual voltage (act), the discharge time of the
performed electroporation, and a voltage symbol appear in the display.
3. Remove cuvette holder from the device.
The cuvette symbol and voltage symbol disappear.
4. Remove electroporation cuvette from the cuvette holder and carefully
transfer the sample to the corresponding medium without bubbles.
5.4 Regeneration of the cells
Example for the bacterium E. coli:
1. After the electroporation, immediately place about 1 mL fresh medium (without
selection chemicals) on the cells. A rich medium is best suited for this, e.g. the SOC
medium for E. coli.
2. Carefully resuspend cells and transfer them to a new tube.
3. Incubate cells at optimal growth temperature (e.g. 37 °C for E. coli) for one hour at
light vibration (e.g. with the Eppendorf Thermomixer comfort).
5.5 Determination of the transformation efficiency
After the recovery period, the cells should be plated with a selection medium.
To determine the efficiency, streak different cell concentrations and use this information
to calculate the number of transformers/μg DNA.
Eppendorf Eporator®
P1
P2
Operation
English (EN)
5.6 Programs
A program contains a saved voltage setting for quick access to frequently used settings.
5.6.1 Loading program
The programs 1 and 2 are stored with the following parameters at delivery:
• Program key P1: 1 700 V
(e.g. for E. coli electroporation in 1 mm electroporation cuvettes)
• Program key P2: 2 500 V
(e.g. for E. coli electroporation in 2 mm electroporation cuvettes)
Press desired program key
The control LED above the pressed program key illuminates blue, the voltage is
displayed.
5.6.2 Saving program
1. Use the arrow keys to set the voltage.
2. Hold down the desired program key for at least 2 seconds.
A signal tone sounds. Voltage stored appears in the display. The control LED
over the pressed program key illuminates blue. The voltage is saved under
the corresponding program number (1 to 2).
19
Operation
1 2
34
menu
enter
menu
enter
exit
Eppendorf Eporator®
20
English (EN)
5.7 Enhanced settings
Additional settings can be carried out in the menu. The date and time can be defined in
the device in order to track exported data. The following settings are available:
Abb. 5-5:Menu display
Fig. 5-5: Menu display
1Data export
The data export is described in the
chapter "Export data" (see p. 21).
2Date
Set date.
Open menu.
1. Press menu/enter key.
Switch between parameters
2. Press menu/enter key.
The selected parameter blinks in the display.
Change the value of the parameter
3. Press the arrow keys.
Exit the menu.
4. Press the exit key.
The changed parameters are automatically saved.
3Time
Set time.
4 Signal tone
Set signal tone. The display switches
between vol1 (very quiet), vol2 (quiet),
vol3 (loud), vol4 (very loud) and vol OFF
(signal tone switched off).
Eppendorf Eporator®
menu
enter
Start
Operation
English (EN)
5.8 Exporting data
Using the USB port at the rear of the device, you can save the last 50 experiments on a
USB stick in separate TXT files (see Main illustration on p. 7). The files are named
according to the corresponding sample number. This format is well-suited for additional
editing in a text editor or Microsoft Excel.
The data sets of an electroporation contain the following information:
• Sample number (sample) of the experiment
For each experiment, the device automatically assigns a four-digit sample number that
is counted from 0001 upwards.
• Date (date) of the experiment
• Time (time) of the experiment
• Set voltage (set) of the experiment
Voltage that was selected for the corresponding experiment using the arrow keys.
• Actual voltage (act) of the experiment
Voltage that was actually applied to the electroporation cuvette in the corresponding
experiment.
• Time constant of the discharge curve (tc) of the experiment
Time constant of the discharge curve of the corresponding experiment.
• Software version (sw) of the device
• Serial number (serial no) of the device
21
Date and time can be specified in the enhanced settings (see p. 20).
Insert USB stick
1. Insert a standard USB stick in the USB port at the rear of the device. (see
Main illustration on p. 7)
Open menu.
2. Press menu/enter key.
The menu is displayed and a cursor blinks in the export display exp OFF.
Activate parameter
3. Press one of the arrow keys.
The selection item exp ON is displayed.
Export data
4. Press the Start key.
The data transfer is started. The main screen is displayed after the export is
completed.
Operation
Eppendorf Eporator®
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English (EN)
Troubleshooting
Eppendorf Eporator®
English (EN)
6 Troubleshooting
6.1 General errors
Many factors can contribute to a low transformation efficiency:
• The set voltage: Specific voltage parameters exist for each microorganism. Some cells
die during electroporation. If the field strength is too high or too low, a low
transformation efficiency is achieved. The expected survival rate varies between 20 %
and 80 % of inserted cells. The electroporation of E. coli requires an impulse of
approximately 5 ms and field strengths between 12 kV/cm and 19 kV/cm. You should
check the transformation efficiency at different voltages in order to optimize
conditions.
Application protocols for the electroporation of various bacteria and yeast strains can
be found on the Eppendorf home page www.eppendorf.com
• The cells: Generally, cells are transformed most efficiently when they are in an early to
medium log phase. Different growth conditions can improve the transformation
efficiency (see Growth and preparation of cells on p. 15).
If too many cells are killed, the electroporation conditions for the strain must be
optimized and the DNA preparation and cell preparation for toxic or organic
substances must be inspected (see DNA preparation on p. 14).
After electroporation, cells (especially E. coli) must be immediately transferred to a rich
medium in order to obtain good results. Even a small delay in completing this step can
lead to a significantly lower transformation efficiency (see Regeneration of the cells on
p. 18).
• The DNA: The quantity and quality of the DNA used should be checked before
electroporation. Incorrectly concentrated or degraded DNA leads to low
transformation efficiency.
Salts and other components that can have a toxic effect on cells must be removed from
the DNA preparation before the preparation process.
The DNA preparation should be added to the cells no longer than one minute before
the electroporation. DNase present in cell preparation can degrade the DNA and
thereby cause a low transformation efficiency (see DNA preparation on p. 14).
• The temperature: Electroporation cuvettes should be cooled to 0 °C - 4 °C before
electroporation (see Temperature on p. 16). This produces better results than with
electroporation cuvettes at room temperature.
If frozen cells are used, electroporation should be performed immediately after
thawing. Frozen cells can be stored a maximum of 6 - 12 months in 10 % - 15 %
glycerine at -80 °C.
.
23
Troubleshooting
Eppendorf Eporator®
24
English (EN)
• Deviating voltages during transformation: The voltage applied to the electroporation
cuvette (act) greatly differs from the set voltage (set)
A too low resistance has several causes:
– The cells were washed and resuspended in a buffer with a too high ionic strength.
– The cells were not sufficiently cleaned during the preparation. After insufficient
washing, growth medium residue, which has been carried along, leaves behind
unwanted salts.
– Lysed cells are in the preparation. They contribute to the reduction of the resistance
of the medium.
– The salt concentration in the DNA preparation is too high.
6.2 Error messages
If the proposed measures repeatedly fail to remedy the fault, contact your local
Eppendorf partner. The addresses of our distributors can be found on our
website www.eppendorf.com
the second to last page of this operating manual.
. The addresses of our sales offices can be found on
Quit all error messages with the Exit key.
6.2.1 Operational error
Troubleshooting
Eppendorf Eporator®
English (EN)
25
Symptom/
message
Display remains
dark
The display
shows:
function not
available
The display
shows:
no cuvette
The display
shows:
no USB stick
The display
shows:
USB stick full
The display
shows:
no export
The display
shows:
no protocol
Cause Remedy
• The device was not connected
to the mains supply or the
mains power switch was not
switched on.
• A key that is not available in the
current device state was
pressed (e.g. the Exit key in the
main display).
•The Start key was pressed
before an electroporation
cuvette was inserted.
• The export command was
activated before a USB stick
was inserted in the USB port of
the device.
• A USB stick with no storage
capacity was inserted in the
USB port of the device.
• The export of the data from the
device has failed.
• All existing electroporation
protocols have already been
saved on the USB stick.
• The export of data has failed.
No exportable protocols are
located in the device.
Check the mains connection
and the power cable.
Switch on the device.
Message disappears after about
2 seconds.
1. Insert an electroporation
cuvette.
2. Start electroporation (see
Inserting the cuvette on p. 16).
1. Insert a USB stick in the USB
port of the device.
2. Repeat the export command
(see Exporting data on p. 21).
1. Insert a USB stick with
sufficient storage capacity in
the USB port of the device.
2. Repeat the export command
(see Exporting data on p. 21).
1. Insert a standard USB stick in
the USB port of the device.
2. Repeat the export command
(see Exporting data on p. 21).
1. Perform electroporation
2. Repeat the export command
(see Exporting data on p. 21).
Troubleshooting
Eppendorf Eporator®
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English (EN)
6.2.2 Device error
Symptom/
message
The display
shows:
Hardware error
The display
shows:
internal error
Cause Remedy
• Device error 1. Quit the error message with the
Exit key.
2. Perform the electroporation
again.
If the error message appears
again: switch the device off and
on.
• Device error 1. Quit the error message with the
Exit key.
2. Perform the electroporation
again.
If the error message appears
again: switch the device off and
on.
Eppendorf Eporator®
1
2
3
Maintenance
English (EN)
7 Maintenance
7.1 Cleaning
DANGER! Electric shock as a result of penetration of liquid.
Switch off the device and disconnect it from the power supply before any
maintenance or cleaning work is carried out.
Do not allow any liquids to enter the inside of the housing.
Do you use any spray disinfectants on the housing.
Only reconnect the device to the power supply once it is completely dry.
NOTICE! Damage from using aggressive chemicals.
Do not use any aggressive chemicals on the device or its accessories, such as
strong and weak bases, strong acids, acetone, formaldehyde, halogenated
hydrocarbons or phenol.
If the device becomes contaminated with aggressive chemicals, clean it
immediately with a mild cleaning agent.
Wet a cloth with a mild cleaning fluid and demineralized water and remove the
contamination on the outside of the device.
7.2 Replacing fuses
The fuse holder is located between the mains/power cord socket and the mains/power
switch.
27
1. Unplug the mains/power plug.
2. Press the upper and lower end of the plastic springs 1 together and pull the fuse holder
2 fully out.
3. Replace faulty fuses and reinsert the fuse holder. Make sure that the guiding rail 3 is
positioned correctly.
Maintenance
Eppendorf Eporator®
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English (EN)
8 Technical data
8.1 Power supply
Mains power connection 100 V to 240 V ± 10 %, 50 Hz to 60 Hz
Automatic adjustment to the voltage
Power consumption: 20 W
Charging time: < 10 s
8.2 Ambient conditions
Environment: For indoor use only
Ambient temperature: 5 °C to 40 °C
Relative humidity: 10 % to 90 %
Atmospheric pressure: 79.5 kPa to 106 kPa (2000 m)
Degree of pollution 2
8.3 Weight/dimensions
Weight: 3.2 kg
Dimensions: Width: 19 cm (7.48 in.)
Height: 12.5 cm (4.92 in.)
Depth: 27.5 cm (10.83 in.)
Technical data
Eppendorf Eporator®
English (EN)
29
8.4 Interfaces
USB: USB 2.0
RS232: Only for Technical Service
8.5 Operating mode
Power-on time: 10 %, 120 s max
Technical data
Eppendorf Eporator®
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English (EN)
9 Ordering Information
9.1 Eporator
Ordering Information
Eppendorf Eporator®
English (EN)
31
Order no.
(International)
4309 000.019 4309 000.027 Eppendorf Eporator
Order no. (North
America)
Description
9.2 Accessories
Order no.
(International)
4308 078.006 940001102 for 16 cuvettes
4309 900.010 4309 900.010 Operating Manual Eppendorf
Order no. (North
America)
Description
Cuvette stand
Eporator
Ordering Information
Eppendorf Eporator®
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English (EN)
Transport, storage and disposal
Eppendorf Eporator®
English (EN)
10 Transport, storage and disposal
10.1 Storage
Air temperature Rel. humidity Air pressure
In transport
packaging
Without transport
packaging
-25 to 55°C 10 to 95% 70 to 106 kPa
-5 to 45°C 10 to 95% 70 to 106 kPa
10.2 Decontamination before shipment
If you are shipping the device to the authorized Technical Service for repairs or to your
authorized dealer for disposal please note the following:
WARNING! Risk to health from contaminated device.
1. Follow the instructions in the decontamination certificate. This can be found
in a PDF file on our homepage (www.eppendorf.com/decontamination)
2. Decontaminate all the parts you want to dispatch.
3. Enclose the fully-completed decontamination certificate for returned goods
(incl. the serial number of the device) with the dispatch.
.
10.3 Transport
Only transport the device in the original packaging.
Air temperature Rel. humidity Air pressure
General
transportation
Air freight -40 °C to 55 °C 10 % to 95 % 30 kPa to 106 kPa
-25 °C to 60 °C 10 % to 95 % 30 kPa to 106 kPa
33
Transport, storage and disposal
Eppendorf Eporator®
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English (EN)
10.4 Disposal
In case the product is to be disposed of, the relevant legal regulations are to be observed.
Information on the disposal of electrical and electronic devices in the European
Community:
Within the European Community, the disposal of electrical devices is regulated by
national regulations based on EU Directive 2002/96/EC pertaining to waste electrical and
electronic equipment (WEEE).
According to these regulations, any devices supplied after August 13, 2005, in the
business-to-business sphere, to which this product is assigned, may no longer be
disposed of in municipal or domestic waste. To document this, they have been marked
with the following identification:
Because disposal regulations may differ from one country to another within the EU,
please contact your supplier if necessary.
In Germany, this is mandatory from March 23, 2006. From this date, the manufacturer has
to offer a suitable method of return for all devices supplied after August 13, 2005. For all
devices supplied before August 13, 2005, the last user is responsible for the correct
disposal.
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Give us your feedback.
www.eppendorf.com/manualfeedback
Your local distributor: www.eppendorf.com/contact
Eppendorf AG · 22331 Hamburg · Germany
eppendorf@eppendorf.com · www.eppendorf.com
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