Bioneer Exicycler 96 Use and care manual

Installation and Calibration Guide
Exicycler™96 Real-Time Quantitative Thermal Block
Exicycler™ 96 Real-Time Quantitative Thermal Block

Warranty and Liability
All Bioneer products are tested under extensive Quality Control procedures. Bioneer guarantees the quality under the
warranty period. Any problems should be reported immediately. Liability is conditional upon the customer providing full
details of the problem to Bioneer. Once the problem occurrs, customers must report to Bioneer within 30 days.
QC Testing
Each lot of Bioneer‟s product is carefully tested by the quality control team.
Notice
Patent Pending
Exicycler is a trademark of Bioneer Corporation.
Certain applications of this product are covered by pending or issued patents in certain countries. Because purchase of
this product does not include a license to perform any patented application, users of this product may be required to
obtain a patent license depending upon the particular application and country in which the product is used. For more
information, please contact Legal manager, Bioneer Corporation, 49-3, Munpyeong-dong, Daedeok-gu, Daejeon 306-220,
Republic of Korea.
Appearance and specification are subjected to change without notice.
Copyright  Feb. 2009 by Bioneer Corporation. All rights reserved. Rev 090815

Contents
Before Starting ....................................................................................................................................4
Safety Warnings and Cautions............................................................................................................9
Location Consideration and Installation ...................................................................9
Operation and Maintenance ................................................................................. 10
European Safety and EMC Standards (CE) ............................................................. 12
Introduction........................................................................................................................................15
System Components and Specifications...........................................................................................19
System Components ........................................................................................... 19
Specifications .................................................................................................... 20
System Views..................................................................................................... 22
Installation.........................................................................................................................................25
Setting Up the Computer ..................................................................................... 25
Connecting the Computer and Exicycler™ 96......................................................... 26
Installing the Operation and Analysis Software........................................................ 27
System Operation and Calibration Preparation.................................................................................35
Powering On and Self-diagnosis .......................................................................... 35
Reagents and Consumable Products..................................................................... 36
Calibration Using ExiCfg....................................................................................................................45
Before Starting Calibration with the Calibration Kit................................................... 45
Calibration Plate Preparation ................................................................................ 45
Calibration using ExiCfg ...................................................................................... 48
Mask Calibration ................................................................................................. 48
Background Calibration ....................................................................................... 51
Multi-channel Calibration..................................................................................... 55
Starting ‘Run Exicycler’ software........................................................................... 63
Performing System Diagnosis using ExiCfg ............................................................ 79
Troubleshooting.................................................................................................................................85
Ordering Information and Warranty...................................................................................................91

Before Starting
How to Use This Guide
Who This Guide is Written for
This guide is written for researchers and laboratory staff responsible for installing and maintaining the
Exicycler™ 96 Real-Time Quantitative Thermal Block.
General Requirements for Installation
This guide assumes that you:
Are familiar with the Microsoft®Windows®XP operating system.
Have basic techniques for handling DNA and RNA samples for PCR.
Have basic skills of data storage, copying, and pasting in hard drives.
Have experience in setting up a network if you want to use any data generated by Exicycler™
96 through a network.
Word Conventions
Bold signifies user action such as typing a text or clicking a button. For example:
Type Test and click OK to move to the next step.
Italic represents important words or sentences and is also used for emphasis. For example:
After analysis, you must save data using Save As.
A right arrow in bold (>) separates consecutive commands you select from a main menu or
shortcut menu. For example:
File> Config> Scan
Safety and Warning
The following safety alert words are used in this manual and require a particular level of observation.
Each alert word is defined as below:
Note: Indicates relevant or helpful information about an instrument, however doesn‟t affect instrument
operation.
Important: Indicates information that is necessary for proper instrument operation.
Danger: Indicates an imminently hazardous situation which, if not avoided, could result in serious injury.
This safety word is used to alert improper instrument operation.
Warning: Indicates a potentially hazardous situation which, if not avoided, will result in death or serious
injury if not avoided.

How to Use an Instrument Safely
Refer to this guide when installing or maintaining the Exicycler™ 96 Real-Time Quantitative Thermal
Block.
How to Obtain More Information
For more information about Exicycler™ 96 , please visit us online at www.bioneer.co.kr or
www.bioneer.com.
How to Reach Customer Support
To obtain prompt customer support, please call us at 1588-9788. You can also obtain technical support
through www.bioneer.co.kr or www.bioneer.com.

Safety Instructions
Exicycler™ 96 Real-Time Quantitative Thermal Block

Exicycler™96 Real-Time Quantitative Thermal Block
9
Safety Warnings and Cautions
Safety instructions in this guide book are to cover possible dangerous conditions and situations that
may occur. It is your responsibility to read this guide thoroughly when installing, maintaining, or
operating Exicycler™ 96. The following safety alert words will be used in this section:
Warning: Hazards or unsafe actions that may result in severe injury.
Caution: Hazards or unsafe actions that may result in minor injury or damage.
Warnings: Hazards or unsafe actions that may result in High temperature and Skin burn.
Warnings: Hazards or unsafe actions that may result in Electronic risk.
Location Consideration and Installation
General Instrument Safety
1) Check the power voltage rating before connecting the instrument to an electrical outlet.
Exicycler™ 96 system is configured for either 110V or 220V (±10%). Using AVR (Automatic
Voltage Regulator) or UPS (Uninterrupted Power Supply) is recommended for uninterruptible
power supply.
2) The Exicycler™ 96 must be grounded for protection against electric shock. If not, it may cause
serious injury and system damage.
3) Do not use a loose power cable or connector.An overheated power cable may break and lead
to a fire or electric shock.
4) If you use a power extension cable, do not connect too many devices to it and operate them at
the same time. Overload may occur and cause a fire.
5) Dry your hands completely when handling a power cord for protection against electric shock.
6) Do not place any objects in front of the main door of the Exicycler™ 96 that can interrupt door
operation.
7) Leave 30 cm space between the Exicycler™ 96 and the wall for proper ventilation.
8) Do not install the Exicycler™ 96 in a dusty environment to help prevent false operation or
technical damage.
9) Keep the Exicycler™ 96 away from heat sources.
10)The Exicycler™ 96 must not be installed in an area where it is exposed to water or is humid. It
may cause electric shock, a fire, or system damage.
11)The Exicycler™ 96 must not be installed in an area where it is exposed to combustible or
flammable vapor. In the case of a gas leak, open the windows and let fresh air in. Do not
operate any electrical switch during a gas leak. It may cause explosion or fire.
12)Do not disassemble or repair the Exicycler™ 96 yourself. It may cause a fire, electric shock,

Exicycler™96 Real-Time Quantitative Thermal Block
10
and system damage. Alimited warranty does not cover unauthorized alterations or damage by
abuse.
Installation Safety
1) Place and install the Exicycler™96 away from direct sunlight.
2) It is recommended that you turn off the computer as well as unplug the power cable before
connecting it to the Exicycler™ 96. When the computer power is on while connecting, the
communication port connector in either the computer or the Exicycler™ 96 may get damaged.
3) Make sure that the USB cable is firmly connected between the Exicycler™ 96 and the
computer. Unstable connection may cause damage of the communication port connector or
data transfer errors.
4) The built-in camera in the Exicycler™ 96 is a static-sensitive device. Pay particular attention to
any cables connected to the Exicycler™ 96 to avoid static damage.
Cautions
1) Ensure the power cable is clean and connect it firmly to the Exicycler™ 96.
2) Operate the Exicycler™96 in a place where the temperature is always between 15℃and 30℃.
Poor performance is influenced by extreme temperatures. High temperatures can cause mis-
operation and poor performance.
3) Operate the system in a place where the humidity is always between 20 % and 80% with no
condensation. High humidity conditions can cause corrosion of internal components and low
humidity can lead to errors.
4) Do not place any objects behind or by the side of the Exicycler™96 that can interrupt
ventilization and cause errors.
5) The internal optical components of the Exicycler™96 may get damaged if the instrument falls
or is exposes to excessive physical shock.
6) Unplug the power cable from the Exicycler™96 when not in operation for a long period of time
to prevent the possibility of fire by overheating.
Operation and Maintenance
Warnings
1) The system can be hazardous if misused.
2) Keep the 96-well thermal block area clean to prevent damage and to generate accurate
experimental data.
3) After any Real-Time PCR runs including „Scan‟ or „Melting‟, allow the Exicycler™ 96 at least 10
minutes to cool down the light source lamp. Continuous operation without a break will reduce
the lamp life time and cause errors.

Exicycler™96 Real-Time Quantitative Thermal Block
11
4) Do not place a piece of paper or a plastic cover under the Exicycler™96. It could cause fire.
5) Do not turn off the Exicycler™96 right after a Real-Time PCR run is done. Wait until the
cooling fan stops running completely. The cooling fan still runs for about 2 minutes to cool
down the lamp after every Real-Time PCR run.
6) Do not cover the Exicycler™96 with a piece of paper or a plastic cover. It may cause a fire or
failure.
7) Set ‟Turn off monitor‟, „Turn off hard disks‟, „System standby„, and „System hibernates‟ to
„Never‟ using „Power Options Properties‟ in the Control Panel. Otherwise, data transfer
between the Exicycler™96 and the computer will be interrupted.
IMPORTANT:
Exicycler™ 96 should be operated in clean condition. Contaminants such as dust can cause troubles and reduce the life
time of Exicycler™ 96. Please prevent dust from coming into Exicycler™ 96. Through removing dust periodically from the
Exicycler™ 96 system, the life time can be extended.

Exicycler™96 Real-Time Quantitative Thermal Block
12
European Safety and EMC Standards (CE)
This instrument meets the requirements of the European Directive for in vitro diagnostic medical
devices 98/79/EC. This instrument has been tested according to the standards listed below and
complies with them.
1) Council Directive 98/79/EC of 27 October 1998 concerning in-vitro diagnostic medical devices
2) EN 61010-1 [2001] Safety requirements for electrical equipment for measurement, control and
laboratory use. Part 1: General requirements for safety.
3) EN 61010-2-010 [2003] Safety requirements for electrical equipment for measurement, control
and laboratory use. Part 2-010: Particular requirements for laboratory equipment for the
heating of materials.
4) EN 61010-2-081 [2001] Safety requirements for electrical equipment for measurement, control
and laboratory use. Part 2-081: Particular requirements for automatic and semi-automatic
laboratory equipment for analysis and other purposes.
5) EN 61010-2-101 [2002] Safety requirements for electrical equipment for measurement, control
and laboratory use. Part 2-101: Particular requirements for in vitro diagnostic (IVD) medical
equipment.
6) EN 61326 [1997+A1] [1998+A2] [2001+A3] [2003 ClassA] Electrical equipment for
measurement, control and laboratory use - EMC requirements.
7) EN 60601-1-2 [2001] Medical electrical equipment. Part 1-2: General requirements for basic
safety and essential performance - Collateral standard: Electromagnetic compatibility -
Requirements and tests.
Compliance is demonstrated by the following mark:
Authorized Representative
MT Promedt Consulting GmbH
Altenhofstrasse 80
D-66386 St. Ingbert
Germany
Product Use Limitations
The Exicycler™system is designed and sold for in vitro diagnostic (IVD) applications in combination
with the diagnostic kits manufactured by Bioneer and labeled for diagnostic purposes. The ExicyclerTM
system can also be used for life science research and other laboratory purposes.

Introduction
Exicycler™ 96 Real-Time Quantitative Thermal Block

Exicycler™ 96 Real Time Quantitative Thermal Block
15
Introduction
The Exicycler™ 96 Real-Time Quantitative Thermal Block combines a thermal block in a standard 96-
well format and an innovative fluorescence detector. The detector monitors the fluorescence emitted as
an indicator of amplified nucleic acid product during each PCR cycle in real time.
The thermal block is built in the lower part of the Exicycler™ 96, which carries out a thermal cycling. The
detector is located in the upper part of the Exicycler™ 96, which measures the fluorescence emitted
from samples in the thermal block in real time and transfers data to the computer for analysis. Data
transferred from the detector is analyzed with the Exicycler ™Analysis Software.
The Exicycler™ 96 Real-Time Quantitative Thermal Block provides a maximum ramping rate of
2.5℃/sec, and features various funtions such as gradient, time increment, temperature increment, and
ramp rate control for myriad applications.
The Exicycler™ 96 Real-Time Quantitative Thermal Block detector consists of an light source part
and a detection part. The light source part is an energy source to excite fluorescent dyes. A short arc
lamp that has a long lifetime is bright, and is implemented. A white-light source is divided into particular
wavelength groups by band path filters. The band path filters that are set in the Exicycler™96 covers
within 480 to 680 ㎚and works as a set consisting of an excitation and an emission filter. Five of the
filter sets are provided for various applications. Therefore, adding additional band path filters is not
required.
The detector, a hight sensitive 2D CCD camera, provides high sensitivity and detects fluorescence
signals from a 96-well plate in the thermal block all at one time. This simultaneous detection has a great
advantage over sequential detection. The innovative detector invented by Bioneer has also reduced
well-to-well variation and has minimized dye-to-dye interference, therefore providing more accurate
results. Bioneer‟s state-of-the-art technologies applied to the Exicycler™ 96 are to generate a uniform
light surface over the thermal block, to detect multiple fluorescent signals emitted from various dyes
through the light surface, and to separate the signals within a selective wavelength range of each dye.
The Exicycler™ 96 Real-Time Quantitative Thermal Block features an auto loading function for
automation reducing errors and Self-diagnosis for diagnosing systematical conditions of the Exicycer™
96 for users convenience. The Exicycler™ 96 Software is composed of three main programs.
1) A set up program for calibration, diagnosis and confirmation of the instrument‟s information.
2) An operation program for creating a protocol, assigning a probe and plate, saving & displaying
data while operating the Exicycer™96.
3) An analysis program for data analysis. The analysis program includes Absolute Quantification,
Relative Quantification, Multi-Relative Quantification, SNP genotyping,
Existentence/Nonexistentence. Analysis and is applicable to Gene expression, Quantification of
cell and virus, and SNP genotyping.

System Components and Specifications
Exicycler™ 96 Real-Time Quantitative Thermal Block

Exicycler™ 96 Real-Time Quantitative Thermal Block
19
System Components and Specifications
Exicycler™ 96 Quantitative Thermal Block is warranted by Bioneer against manufacturing defects in
materials and workmanship for a limited warranty period of one year. Bioneer will charge for repairing
products for the following conditions:
Failures due to work by the customer himself/herself
The product is repaired after expiry of the warranty period.
Before you install the Exicycler™ 96, check the shipped materials with the system component list below:
System Components
Cat. No. A-2060
Check
Main instrument
1
□
USB 2.0 high speed cable
1
□
Power cable
1
□
Software
1 CD
□
Operations Guide
1
□
96 well reaction plate
1 pack
□
Optical sealing tape
1 pack
□
Optical tape sealing applicator
1
□
Warranties
1
□
Customer Service Request Form
1
□

Exicycler™ 96 Real-Time Quantitative Thermal Block
20
Specifications
Physical specifications
Dimension (㎜)
355(W) X 540(D) X 470(H)
Weight (㎏)
30 ㎏
Sample capacity size
96-well plate / 0.2 ㎖micro tubes
Sample volume
20~50 ㎕
(50 ㎕recommended)
Input voltage
100 ~ 240 VAC
Frequency
50 / 60 ㎐
Power
850 Watts (Fuse : 250 V, F10AL)
Operating specifications
Method of heating / cooling
Peltier
Temperature range
4.0℃~ 99.9℃
Temperature accuracy
0.3℃
Temperature uniformity
0.5℃
Ramping rate
Max. 2.5℃/sec
Lid temperature
Within 90 ~ 120℃
Gradient range
1℃~20℃
Temperature increment range
0.1℃~9.9℃
Time increment range
1 sec ~ 60 sec
Ramp rate control
0.1℃/sec
Operating temperature
15 ~ 30℃
Operating humidity
20 ~ 80%, no condensation
Communication
USB 2.0 high speed
Operating OS
Windows XP (either in English or in Korean)
Optical Part
Light source
Short arc lamp
Wattage
120 W
* Lamp life time
3,000 hours
Sensor
16 bit 2D CCD
** Excitation Filter / Emission Filter
5 Types / 5 Types
* Continuously turning on and off the lamp reduces the lamp life time.
* The lamp generates heat when it starts. Make sure that you turn off the Exicycler™ after a cooling fan
stops running. It usually takes about 2 minutes to stop the cooling fan. Please refer to a chart below
for more information about filter sets.

Exicycler™ 96 Real-Time Quantitative Thermal Block
21
† Filter sets
Warning:
Chemical hazard: Such the florescent dyes may cause eye and skin irritation, and respiratory tract irritation. To treat
them, read the MSDS before use and follow the instructions if swallowed or inhaled. Wear appropriate protective
eyewear, clothing, and gloves.
Position
Excitation (㎚)
Emission (㎚)
Set
Fluorescent dye
1
Blank
Blank
1
-
2
490
520
1
FAM, SYBR Green I
3
520
550
1
JOE, TET
4
550
580
1
TAMRA, CY3
5
580
610
1
Texas Red, ROX, Red610
6
640
670
1
CY5, Red670

Exicycler™ 96 Real-Time Quantitative Thermal Block
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System Views
Front view Rear view
Inside view
96-well thermal block
USB Communication cable
On/Off Switch
DOOR
POWER and Status LED
Ventilation hole

Installation
Exicycler™ 96 Real-Time Quantitative Thermal Block

Exicycler™ 96 Real-Time Quantitative Thermal Block
25
Installation
To run the Exicycler™ 96 Real-Time Quantitative Thermal Block, camera driver and operation software
should be installed in a computer. When first installing the camera driver and operation software, or re-
installing these components, please see below for instructions on how to install them yourself.
Setting Up the Computer
Exicycler™ 96 Real-Time Quantitative Thermal Block operates through a computer. Therefore, to install
Exicycler™ 96, a computer or a laptop is necessary. The computer must meet the following computer
requirements listed below. Please contact Bioneer Customer Service Center if you have any questions
regarding installation.
Computer Requirements
Intel Dual Core E2160 (1.8GHz) or higher
Window XP Operating System for Korean or English Version (Service pack 2 or later)
1.0GB RAM or higher
1280 * 1024 screen resolution
USB 2.0 high speed port
20GB Hard disk drive minimum
Microsoft Excel (Option)
For questions regarding problems with the computer or operating system, please contact the computer
manufacturer.
1) The computer must have at least one communication port for USB data transfer.
2) It is recommended that antivirus software be installed in order for the Exicycler™96 to operate
safely. A firewall should also be setup to prevent unwanted information coming in from external
nextworks. Please contact your IT department to setup the antivirus software and firewall.
3) It is recommended that a memory stick not be installed when Exicycler™96 is running. It may cause
a technical problem between Exicycler™96 and the computer.

Exicycler™ 96 Real-Time Quantitative Thermal Block
26
Connecting the Computer and Exicycler™ 96
There are a main power connector and a communication port connector (USB) in the rear of Exicycler™
96. The computer should be turned off when connecting to the Exicycler™96 to prevent any damages
to the communication port. Please see below for instructions on how to connect Exicycler™96 to the
computer.
1) Unpack the Exicycler™ 96 and make sure all components are included. Refer to the list of
components below.
2) Exicycler™96 must be installed in an area where it is not exposed to sunlight and must be set on a
stable and level surface.
3) Set up the computer in the installation site.
4) Place the Exicycler™96 in the installation site carefully.
5) Connect the USB 2.0 cable to the USB connector on the back of Exicycler™ 96, then to the
computer.
6) Connect the power cable to the Exicycler™ 96, then to the receptable wall circuit.
List of Components
* Provided according to country standard.
Cat. No. A-2060
Check
Main instrument
1
□
USB 2.0 high speed cable
1
□
Power cable*
1
□
Software
1 CD
□
Operations Guide
1
□
96 well reaction plate
1 pack
□
Optical sealing tape
1 pack
□
Optical tape sealing applicator
1
□
Warranties
1
□
Customer Service Request Form
1
□