Eporator
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nualorator®
E
N)
manual
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Eppendorf Eporator®
Operating manual
4309 900.010-04/082013
Copyright
©
2013 Eppendorf AG, Hamburg. No part of this publication may be reproduced
without the prior permission of the copyright owner.
Eppendorf
®
and the Eppendorf logo are registered trademarks of Eppendorf AG,
Hamburg, Germany.
Eppendorf Eporator
®
is a registered trademark of Eppendorf AG, Hamburg, Germany.
Excel
®
and Microsoft
®
are registered trademarks of Microsoft Corporation in the United
States and other countries.
Registered trademarks are not marked in all cases with ™ or ® in this manual.
Table of contents
Eppendorf Eporator®
English (EN)
3
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
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®
English (EN)
4
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)
5
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
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.
1.3 Symbols used
Toxic substances Electric shock
Hazard point Material damage
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.
Representation Meaning
Handling
1.
2.
Actions in the specified order
• List:
Text Name of fields in the software
Useful information
Operating instructions
Eppendorf Eporator®
English (EN)
6
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.
Product description
Eppendorf Eporator®
English (EN)
7
2 Product description
2.1 Main illustration
Abb. 2-1:Main Illustration
Fig. 2-1: Main Illustration
2.2 Delivery package
1 Operating controls
2 Cuvette carrier
In the cuvette shaft
3 RS232 port
Only for Technical Service
4USB port
For inserting a USB stick
5Mains switch
6 Mains connection socket
Quantity Description
1 Eppendorf Eporator
1Mains/power cord
1 Cuvette holder
1 Eporator operating manual
1 2
3 4
56
Product description
Eppendorf Eporator®
English (EN)
8
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
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 and bacteria Electrical charge
200 V - 2 500 V;
approx. 5 ms
DNA in bacteria
Safety
Eppendorf Eporator®
English (EN)
9
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
3.5 Warnings for intended use
Depiction Meaning
WAR NING
Follow the operating manual.
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®
English (EN)
10
WARNING! Lethal voltages inside the device.
Ensure that the housing is always closed and undamaged so that no parts
inside the device can be contacted by accident.
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
have been specifically authorized by Eppendorf.
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
undamaged.
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
electrical requirements on the name plate.
Only use sockets with a protective grounding conductor and a suitable mains
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.
Installation
Eppendorf Eporator®
English (EN)
11
4 Installation
4.1 Preparing installation
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
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.
Retain the transport carton and packing material for subsequent safe transport
or storage.
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.
Installation
Eppendorf Eporator®
English (EN)
12
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