orator® nual gEN) manual Register your instrument! www.eppendorf.
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® 3 English (EN) Table of contents 1 Operating instructions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2.1 Hazard symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of contents 4 Eppendorf Eporator® English (EN) 6 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1 General errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2 Error messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.1 Operational error . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.
Operating instructions Eppendorf Eporator® 5 English (EN) 1 1.1 Operating instructions 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.
Operating instructions 6 Eppendorf Eporator® 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.
Product description Eppendorf Eporator® 7 English (EN) 2 2.1 Product description Main illustration Abb. 2-1:Main Illustration 1 2 3 4 6 5 Fig. 2-1: Main Illustration 1 Operating controls 4 2 Cuvette carrier In the cuvette shaft 5 Mains switch 3 RS232 port Only for Technical Service 6 Mains connection socket 2.
Product description 8 Eppendorf Eporator® 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.
Safety Eppendorf Eporator® 9 English (EN) 3 3.1 Safety 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.
Safety 10 Eppendorf Eporator® English (EN) 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.
Installation Eppendorf Eporator® 11 English (EN) 4 4.1 Installation 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.
Installation 12 Eppendorf Eporator® English (EN)
Operation Eppendorf Eporator® 13 English (EN) 5 5.1 Operation 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 1 2 P2 P1 6 menu enter 5 exit Start 3 4 Fig. 5-1: 1 2 3 Operating controls 4 P1 program key with control LED Press: load voltage value. Press and hold (> 2 s): save current voltage value. 5 P2 program key with control LED Press: load voltage value.
Operation 14 Eppendorf Eporator® English (EN) Abb. 5-2:Display 2 1 5 Fig. 5-2: 4 4 1 Actual voltage value (in V) 2 Actual discharge time (in ms) 3 Voltage symbol 5 The voltage symbol appears after electroporation and disappears after the cuvette carrier has been removed. 5.2 3 Display Cuvette symbol The cuvette symbol cuvette is inserted.
Operation Eppendorf Eporator® 15 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.
Operation 16 Eppendorf Eporator® 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 5.3.1 Performing electroporation Switching on the device Press the mains power switch on the rear of the device to switch on the device. 5.3.
Operation Eppendorf Eporator® 17 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 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 18 Eppendorf Eporator® 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). Start 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.
Operation Eppendorf Eporator® 19 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.
Operation 20 Eppendorf Eporator® 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 1 2 4 Fig. 5-5: 3 Menu display 1 Data export The data export is described in the chapter "Export data" (see p. 21). 2 Date Set date. 3 Time Set time. 4 Signal tone Set signal tone.
Operation Eppendorf Eporator® 21 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.
Operation 22 Eppendorf Eporator® English (EN)
Troubleshooting Eppendorf Eporator® 23 English (EN) 6 6.1 Troubleshooting 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.
Troubleshooting 24 Eppendorf Eporator® 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.
Troubleshooting Eppendorf Eporator® 25 English (EN) 6.2.1 Operational error Symptom/ message Cause Display remains • The device was not connected dark to the mains supply or the mains power switch was not switched on. Remedy Check the mains connection and the power cable. Switch on the device. The display shows: function not available • A key that is not available in the Message disappears after about current device state was 2 seconds. pressed (e.g. the Exit key in the main display).
Troubleshooting 26 Eppendorf Eporator® English (EN) 6.2.2 Device error Symptom/ message Cause Remedy The display shows: Hardware error • 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. The display shows: internal error • Device error 1. Quit the error message with the Exit key. 2. Perform the electroporation again.
Maintenance Eppendorf Eporator® 27 English (EN) 7 7.1 Maintenance 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.
Maintenance 28 Eppendorf Eporator® English (EN)
Technical data Eppendorf Eporator® 29 English (EN) 8 8.1 Technical data 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.
Technical data 30 Eppendorf Eporator® English (EN)
Ordering Information Eppendorf Eporator® 31 English (EN) 9 9.1 Ordering Information Eporator Order no. (International) 4309 000.019 Order no. (North America) 4309 000.027 Description Order no. (International) Order no. (North America) Description 4308 078.006 4309 900.010 940001102 4309 900.010 9.
Ordering Information 32 Eppendorf Eporator® English (EN)
Transport, storage and disposal Eppendorf Eporator® 33 English (EN) 10 10.1 Transport, storage and disposal Storage Air temperature Rel. humidity Air pressure In transport packaging -25 to 55°C 10 to 95% 70 to 106 kPa Without transport packaging -5 to 45°C 10 to 95% 70 to 106 kPa 10.
Transport, storage and disposal 34 Eppendorf Eporator® 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).
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