Qiagen Rotor-Gene Q User manual
November 2012
Rotor-Gene® Q User Manual
Sample & Assay Technologies
Trademarks
®
QIAGEN
, QIAgility®, EpiTect®, HotStarTaq®, HRM®, Quantiscript®, QuantiTect®, Rotor-Gene®, Rotor-Disc®, Type-it® (QIAGEN Group); CAL Fluor®, Quasar® (Biosearch Technologies, Inc.); Cy®
(GE Healthcare); EvaGreen® (Biotium, Inc.); LC Green® (Idaho Technology, Inc.); Alexa Fluor®, FAM™, HEX™, JOE™, Marina Blue®, ROX™, SYBR®, SYTO®, TET™, Texas Red®, VIC® (Life Technologies
Corporation); Yakima Yellow® (Nanogen, Inc.); LightCycler® (Roche Group); Core™, Intel® (Intel Corporation); Adobe®, Illustrator® (Adobe Systems, Inc.); Microsoft®, Windows®, Excel® (Microsoft
Corporation). Registered names, trademarks, etc. used in this document, even when not specifically marked as such, are not to be considered unprotected by law.
TeeChartOffice: Copyright 2001-2002 by David Berneda. All rights reserved.
For applicable countries:
This real-time thermal cycler is licensed under pending U.S. Patent rights for an apparatus or system covering automated thermal cyclers with fluorescence detectors and seeking priority to U.S.
Serial No. 07/695,201 and corresponding claims in any foreign counterpart patent thereof owned by Applied Biosystems LLC, in all fields, including research and development, all applied fields,
and human and animal in-vitro diagnostics. No rights are conveyed expressly, by implication or estoppel to any patents on real-time methods, including but not limited to 5' nuclease assays, or to any
patent claiming a reagent or kit. For further information on purchasing additional rights, contact the Director of Licensing at Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California,
94404, USA.
For applicable countries:
The purchase of this product includes a limited, non-transferable license to one or more of US Patents Nos 6,787,338; 7,238,321; 7,081,226; 6,174,670; 6,245,514; 6,569,627; 6,303,305;
6,503,720; 5,871,908; 6,691,041; 7,387,887; and U.S. Patent Applications Nos. 2003-0224434 and 2006-0019253 and all continuations and divisionals, and corresponding claims in
patents and patent applications outside the United States, owned by the University of Utah Research Foundation, Idaho Technology, Inc., and/or Roche Diagnostics GmbH, for internal research
use or for non-in vitro diagnostics applications. No right is conveyed, expressly, by implication or estoppel, for any reagent or kit, or under any other patent or patent claims owned by the
University of Utah Research Foundation, Idaho Technology, Inc., and/or Roche Diagnostics GmbH, or by any other Party. For information on purchasing licences for in-vitro diagnostics applications
or reagents, contact Roche Molecular Systems, 4300 Hacienda Drive, Pleasanton, CA 94588, USA.
For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at
www.qiagen.com or can be requested from QIAGEN Technical services or your local distributors.
© 2005–2012 QIAGEN, all rights reserved.
Contents
Contents
1 Safety Information 1-1
1.1 Proper use 1-2
1.2 Electrical safety 1-4
1.3 Environment 1-5
1.4 Biological safety 1-5
1.5 Chemicals 1-6
1.6 Waste disposal 1-7
1.7 Mechanical hazards 1-7
1.8 Heat hazard 1-8
1.9 Maintenance 1-9
1.10 Symbols on the Rotor-Gene Q 1-10
2 Introduction 2-1
2.1 About this user manual 2-1
2.2 General Information 2-2
2.2.1 Technical assistance 2-2
2.2.2 Policy statement 2-2
2.2.3 Version management 2-2
2.3 Intended use of the Rotor-Gene Q 2-3
3 General Description 3-1
3.1 Thermal performance 3-1
3.2 Optical system 3-3
4 Installation Procedures 4-1
4.1 Site requirements 4-1
4.2 AC Power connection 4-2
Rotor-Gene Q User Manual 11/2012 Contents-1
Contents
4.3 PC requirements 4-2
4.4 Unpacking the Rotor-Gene Q 4-3
4.5 Accessories 4-4
4.6 Hardware installation 4-4
4.7 Software installation 4-6
4.8 Software version 4-9
4.9 Additional software on connected computers 4-10
4.9.1 Virus scanners 4-11
4.9.2 System tools 4-11
4.9.3 Operating system updates 4-12
4.10 Updating software 4-12
5 Operating Procedures — Hardware 5-1
5.1 Rotor types 5-1
5.2 Reaction setup 5-4
5.3 Rotor-Disc setup 5-9
6 Operating Procedures — Software 6-1
6.1 Quick Start wizard 6-1
6.1.1 Rotor selection 6-4
6.1.2 Confirm profile 6-5
6.1.3 Save run 6-6
6.1.4 Sample setup 6-7
6.2 Advanced wizard 6-7
6.2.1 New Run Wizard window 1 6-9
6.2.2 New Run Wizard window 2 6-10
6.2.3 New Run Wizard window 3 6-11
6.2.4 Edit Profile 6-12
6.2.5 New Run Wizard window 4 6-31
6.2.6 New Run Wizard window 5 6-31
Contents-2 Rotor-Gene Q User Manual 11/2012
Contents
7 Analysis User Interface 7-1
7.1 Workspace 7-1
7.2 Toolbar 7-1
7.3 View raw channels 7-1
7.4 Toggling samples 7-3
7.5 File menu 7-5
7.5.1 New 7-5
7.5.2 Open and Save 7-7
7.5.3 Reports 7-9
7.5.4 Setup 7-9
7.6 Analysis menu 7-11
7.6.1 Analysis 7-11
7.6.2 Quantitation 7-12
7.6.3 Two standard curve 7-31
7.6.4 Delta delta CT relative quantitation 7-36
7.6.5 Melt curve analysis 7-40
7.6.6 Comparative quantitation 7-44
7.6.7 Allelic discrimination 7-47
7.6.8 Scatter graph analysis 7-49
7.6.9 EndPoint analysis 7-52
7.6.10 Concentration analysis 7-61
7.6.11 High Resolution Melt analysis 7-64
7.7 Run menu 7-65
7.7.1 Start Run 7-65
7.7.2 Pause Run 7-66
7.7.3 Stop Run 7-66
7.8 View menu 7-66
7.8.1 Run Settings 7-66
7.8.2 Temperature Graph 7-71
7.8.3 Profile Progress 7-72
7.8.4 Edit Samples 7-73
7.8.5 Display Options 7-83
Rotor-Gene Q User Manual 11/2012 Contents-3
Contents
7.9 Security menu 7-84
7.9.1 Configuration 7-85
7.9.2 Running multiple users on the same computer 7-95
7.9.3 Audit trails 7-96
7.9.4 Run Signatures 7-98
7.9.5 Sample locking 7-100
7.9.6 Locked templates 7-102
7.10 Gain menu 7-103
7.11 Window menu 7-104
7.12 Help function 7-104
7.12.1 Send Support E-Mail 7-104
8 Additional Functions 8-1
8.1 Analysis templates 8-1
8.2 Opening a second run 8-1
8.3 Scaling options 8-1
8.4 Exporting graphs 8-2
8.5 Spanner/wrench icon 8-5
8.6 Selected area options 8-7
9 Maintenance Procedures 9-1
10 Optical Temperature Verification 10-1
10.1 OTV principle 10-1
10.2 Rotor-Disc OTV Kit components 10-2
10.3 Running an OTV 10-2
11 High Resolution Melt Analysis 11-1
11.1 Instrumentation 11-3
11.2 Chemistry 11-3
Contents-4 Rotor-Gene Q User Manual 11/2012
Contents
11.3 SNP genotyping example 11-3
11.4 Methylation analysis example 11-5
11.5 Guidelines for successful HRM analysis 11-7
11.6 Sample preparation 11-9
11.7 Software setup 11-9
11.8 Real-time PCR data analysis 11-17
11.9 HRM data analysis 11-19
12 Troubleshooting 12-1
12.1 Log Archives 12-1
12.2 HRM troubleshooting 12-1
12.3 General instrument errors 12-3
13 Glossary 13-1
Appendix A A-1
Technical data A-1
Environmental conditions A-1
FCC Declaration A-4
Declaration of Conformity A-6
Waste Electrical and Electronic Equipment (WEEE) A-7
Appendix B B-1
Safety Information (French, FR) B-1
1 Informations de sécurité B-1
1.1 Utilisation appropriée B-2
1.2 Sécurité électrique B-4
1.3 Environnement B-6
Rotor-Gene Q User Manual 11/2012 Contents-5
Contents
1.4 Sécurité biologique B-6
1.5 Produits chimiques B-8
1.6 Mise au rebut des déchets B-8
1.7 Dangers mécaniques B-9
1.8 Danger lié à la chaleur B-10
1.9 Maintenance B-11
1.10 Symboles du Rotor-Gene Q B-12
Appendix C C-1
Safety Information (German, DE) C-1
1 Sicherheitshinweise C-1
1.1 Sachgemäße Handhabung C-2
1.2 Schutz vor Stromschlag C-4
1.3 Umgebungsbedingungen C-6
1.4 Biologische Sicherheit C-6
1.5 Chemikalien C-8
1.6 Entsorgen von Abfällen C-8
1.7 Gefahren durch mechanische Teile C-9
1.8 Überhitzungsgefahr C-10
1.9 Wartungsarbeiten C-11
1.10 Symbole auf dem Rotor-Gene Q C-12
Appendix D D-1
Quantitation D-1
Appendix E E-1
Rotor-Gene Q products, accessories, and consumables E-1
Contents-6 Rotor-Gene Q User Manual 11/2012
Contents
Appendix F F-1
Liability clause F-1
Index Index-1
Rotor-Gene Q User Manual 11/2012 Contents-7
Contents
This page intentionally left blank
Contents-8 Rotor-Gene Q User Manual 11/2012
1 Safety Information
this one.
Before using the Rotor-Gene Q, it is essential that you read
this user manual carefully and pay particular attention to the
safety information. The instructions and safety information in
the user manual must be followed to ensure safe operation
of the instrument and to maintain the instrument in a safe
condition.
Note: Translations in French and German are available in
Appendix B and Appendix C.
The following types of safety information appear throughout
WARNING
CAUTION
this manual.
The term WARNING is used to inform you about situations
that could result in personal injury to you or other
persons.
Details about these circumstances are given in a box like
this one.
The term CAUTION is used to inform you about situations
that could result in damage to the instrument or other
equipment.
Details about these circumstances are given in a box like
Safety Information
The advice given in this manual is intended to supplement,
not supersede, the normal safety requirements prevailing in
the user’s country.
Rotor-Gene Q User Manual 11/2012 1-1
Safety Information
1.1 Proper use
WARNING/
CAUTION
WARNING/
CAUTION
WARNING/
CAUTION
Risk of personal injury and material damage [W1]
Improper use of the Rotor-Gene Q may cause personal
injuries or damage to the instrument.
The Rotor-Gene Q must only be operated by qualified
personnel who have been appropriately trained.
Servicing of the Rotor-Gene Q must only be performed by
QIAGEN Field Service Specialists.
Perform the maintenance as described in Section 9. QIAGEN
charges for repairs that are required due to incorrect
maintenance.
Risk of personal injury and material damage [W2]
Rotor-Gene Q is a heavy instrument. To avoid personal
injury or damage to the instrument, take care when lifting.
Risk of personal injury and material damage [W3]
Do not attempt to move the Rotor-Gene Q during
operation.
CAUTION
1-2 Rotor-Gene Q User Manual 11/2012
Damage to the instrument [C1]
Avoid spilling water or chemicals onto the Rotor-Gene Q.
Damage caused by water or chemical spillage will void
your warranty.
Note: In case of emergency, switch off the Rotor-Gene Q at
the power switch at the back of the instrument and unplug
the power cord from the power outlet.
Note: Do not switch off the instrument during a run except
for reasons mentioned in this manual. Powering off during a
run has incalculable effects on sample and analysis results. It
is at your own risk to continue a run after it has been
interrupted by powering off the instrument.
WARNING/
CAUTION
WARNING/
CAUTION
WARNING/
CAUTION
WARNING/
CAUTION
CAUTION
CAUTION
Safety Information
Risk of personal injury and material damage
Do not try to open the lid during an experiment, or while
the Rotor-Gene Q is spinning. Otherwise, if you overcome
the lid lock and reach inside, you risk contact with parts
that are hot, electrically live, or moving at high speed, and
you may injure yourself and damage the instrument.
Risk of personal injury and material damage
If you need to stop an experiment quickly, turn off the
power to the instrument, then open the lid. Let the
chamber cool before reaching inside. Otherwise you risk
injury by touching parts that are hot.
Risk of personal injury and material damage [W6]
If the equipment is used in a manner not specified by the
manufacturer, the protection provided by the equipment
may be impaired.
Risk of personal injury and material damage [W7]
Loose paper underneath the Rotor-Gene Q interferes with
instrument cooling. It is recommended that the area
beneath the instrument is kept free of clutter.
Damage to the instrument
Always use a locking ring on the rotor. This stops caps
from coming off tubes during an experiment. If caps come
off during an experiment, they may damage the chamber.
Damage to the instrument [C3]
Visually inspect and make sure the rotor is not damaged or
deformed before each run.
If you touch the Rotor-Gene Q during an experiment, while
you are charged with static electricity, in severe cases the
Rotor-Gene Q may reset. However, the software will restart
the Rotor-Gene Q and continue the experiment.
[W4]
[W5]
[C2]
Rotor-Gene Q User Manual 11/2012 1-3
Safety Information
operated under these conditions.
1.2 Electrical safety
Disconnect the line power cord from the power outlet before
servicing.
WARNING
Electrical hazard [W8]
Any interruption of the protective conductor (earth/ground
lead) inside or outside the instrument or disconnection of
the protective conductor terminal is likely to make the
instrument dangerous.
Intentional interruption is prohibited.
Lethal voltages inside the instrument
When the instrument is connected to line power, terminals
may be live, and opening covers or removing parts is likely
to expose live parts.
To ensure satisfactory and safe operation of the Rotor-Gene
Q, follow the advice below:
The line power cord must be connected to a line power
outlet that has a protective conductor (earth/ground).
Do not adjust or replace internal parts of the instrument.
Do not operate the instrument with any covers or parts
removed.
If liquid has spilled inside the instrument, switch off the
instrument, disconnect it from the power outlet, and
contact QIAGEN Technical Services.
If the instrument becomes electrically unsafe, prevent other
personnel from operating it, and contact QIAGEN Technical
Services; the instrument may be electrically unsafe when:
It or the line power cord appears to be damaged.
It has been stored under unfavorable conditions for a
prolonged period.
It has been subjected to severe transport stresses.
WARNING
Electrical hazard
The instrument has an electrical compliance label which
indicates the voltage and frequency of the power supply as
well as fuse ratings. The equipment should only be
1-4 Rotor-Gene Q User Manual 11/2012
[W9]
1.3 Environment
CAUTION
Damage to the instrument
Operating conditions
WARNING
Explosive atmosphere [W10]
The Rotor-Gene Q is not designed for use in an explosive
atmosphere.
Direct sunlight may bleach parts of the instrument and
cause damage to plastic parts.
The Rotor-Gene Q must be located out of direct sunlight.
1.4 Biological safety
Specimens and reagents containing materials from
biological sources should be treated as potentially infectious.
Use safe laboratory procedures as outlined in publications
such as Biosafety in Microbiological and Biomedical
Laboratories, HHS (www.cdc.gov/od/ohs/biosfty/biosfty.htm).
Safety Information
[C4]
Rotor-Gene Q User Manual 11/2012 1-5
Samples
Samples may contain infectious agents. You should be aware
of the health hazard presented by such agents and should
use, store, and dispose of such samples according to the
required safety regulations.
Safety Information
safety regulations and laws.
WARNING
Samples containing infectious agents [W11]
Some samples used with this instrument may contain
infectious agents. Handle such samples with the greatest of
care and in accordance with the required safety
regulations.
Always wear safety glasses, 2 pairs of gloves, and a lab
coat.
The responsible body (e.g., laboratory manager) must take
the necessary precautions to ensure that the surrounding
workplace is safe, and that the instrument operators are
suitably trained and not exposed to hazardous levels of
infectious agents as defined in the applicable Material
Safety Data Sheets (MSDSs) or OSHA,* ACGIH,
‡
COSHH
documents.
Venting for fumes and disposal of wastes must be in
accordance with all national, state, and local health and
safety regulations and laws.
1.5 Chemicals
WARNING
Hazardous chemicals [W12]
Some chemicals used with this instrument may be
hazardous or may become hazardous after completion of
the protocol run.
Always wear safety glasses, gloves, and a lab coat.
The responsible body (e.g., laboratory manager) must take
the necessary precautions to ensure that the surrounding
workplace is safe and that the instrument operators are not
exposed to hazardous levels of toxic substances (chemical
or biological) as defined in the applicable Material Safety
Data Sheets (MSDSs) or OSHA,* ACGIH,
documents.
Venting for fumes and disposal of wastes must be in
accordance with all national, state, and local health and
†
or
†
or COSHH‡
* OSHA: Occupational Safety and Health Administration (United States of America).
†
ACGIH: American Conference of Government Industrial Hygienists (United States of America).
‡
COSHH: Control of Substances Hazardous to Health (United Kingdom).
1-6 Rotor-Gene Q User Manual 11/2012
Toxic fumes
If working with volatile solvents or toxic substances, you must
provide an efficient laboratory ventilation system to remove
vapors that may be produced.
1.6 Waste disposal
Used consumables and plasticware may contain hazardous
chemicals or infectious agents. Such wastes must be collected
and disposed of properly according to local safety
regulations.
1.7 Mechanical hazards
The lid of the Rotor-Gene Q must remain closed during
WARNING
WARNING/
CAUTION
operation of the instrument.
Moving parts [W13]
To avoid contact with moving parts during operation of the
Rotor-Gene Q, the instrument must be operated with the
lid closed.
Risk of personal injury and material damage [W14]
Open and close the lid of the Rotor-Gene Q carefully to
avoid trapping fingers or clothing.
Safety Information
CAUTION
Rotor-Gene Q User Manual 11/2012 1-7
Damage to the instrument [C5]
Make sure that the rotor and locking ring are installed
correctly.
If the rotor or locking ring show signs of mechanical
damage or corrosion, do not use the Rotor-Gene Q;
contact QIAGEN Technical Services.
Safety Information
WARNING
Hot surface
CAUTION
CAUTION
WARNING
WARNING
Damage to the instrument [C6]
The Rotor-Gene Q must not be used if the lid is broken or
if the lid lock is damaged.
Make sure that the rotor and locking ring are installed
correctly.
Only use rotors, locking rings, and consumables designed
for use with the Rotor-Gene Q. Damage caused by use of
other consumables will void your warranty.
Damage to the instrument [C7]
When Rotor-Gene Q is started immediately after delivery
in cold climates, mechanical parts can block.
Allow the instrument to acclimatize to room temperature
for at least an hour before turning the instrument on.
Moving parts [W15]
In case of breakdown caused by power failure, remove the
power cord and wait 10 minutes before attempting to
manually open the lid.
Risk of overheating [W16]
To ensure proper ventilation, maintain a minimum
clearance of 10 cm at the sides and rear of the
Rotor-Gene Q.
Slits and openings that ensure the ventilation of the
Rotor-Gene Q must not be covered.
1.8 Heat hazard
1-8 Rotor-Gene Q User Manual 11/2012
[W17]
The Rotor-Gene Q chamber can reach temperatures
above 120°C (248°F). Avoid touching it when it is hot.
Safety Information
WARNING
Hot surface [W18]
When a run is paused, the Rotor-Gene Q will not be
cooled completely to room temperature. Exercise caution
before handling the rotor or any tubes in the instrument.
1.9 Maintenance
Perform the maintenance as described in Section 9. QIAGEN
charges for repairs that are required due to incorrect
WARNING/
CAUTION
WARNING
WARNING/
CAUTION
maintenance.
Risk of personal injury and material damage [W19]
Only perform maintenance that is specifically described in
this user manual.
Risk of fire [W20]
When cleaning the Rotor-Gene Q with alcohol-based
disinfectant, leave the Rotor-Gene Q lid open to allow
flammable vapors to disperse.
Only clean the Rotor-Gene Q when the chamber has
cooled down.
Risk of electrical shock [W21]
Do not disassemble the Rotor-Gene Q instrument.
CAUTION
Rotor-Gene Q User Manual 11/2012 1-9
Damage to the instrument housing [C8]
Never clean the instrument housing with alcohol or
alcohol-based solutions. Alcohol will damage the housing.
To clean the housing, use distilled water only.
Safety Information
the temperature
Type plate on the back
Type plate on the back
Type plate on the back
Type plate on the back
Type plate on the back
Type plate on the back
Type plate on the back
1.10 Symbols on the Rotor-Gene Q
Symbol Location Description
Near the sample
chamber, visible when
lid is open
Back of the instrument Consult instructions for use
of the instrument
of the instrument
Heat hazard —
of the chamber can reach
temperatures above 120°C
(248°F)
CE marking for European
Conformity
CSA listing mark for Canada
and the USA
1-10 Rotor-Gene Q User Manual 11/2012
of the instrument
of the instrument
of the instrument
of the instrument
of the instrument
Legal manufacturer
Waste Electrical and Electronic
Equipment (WEEE)
FCC mark of the United States
Federal Communications
Commission
C-Tick mark for Australia
(supplier identification N17965)
RoHS mark for China (the
restriction of the use of certain
hazardous substances in
electrical and electronic
equipment)
2 Introduction
Thank you for choosing the Rotor-Gene Q. We are confident
it will become an integral part of your laboratory.
Before using the Rotor-Gene Q, it is essential that you read
this user manual carefully and pay particular attention to the
safety information. The instructions and safety information in
the user manual must be followed to ensure safe operation
of the instrument and to maintain the instrument in a safe
condition.
2.1 About this user manual
This user manual provides information about the
Rotor-Gene Q in the following sections:
1. Safety Information
2. Introduction
3. General Description
4. Installation Procedures
5. Operating Procedures — Hardware
6. Operating Procedures — Software
7. Analysis User Interface
8. Additional Functions
9. Maintenance Procedures
10. Optical Temperature Verification
11. High Resolution Melt Analysis
12. Troubleshooting
13. Glossary
The appendices contain the following:
Technical data
Safety information in French and German
Mathematical techniques
Declaration of Conformity
Rotor-Gene Q accessories
Liability clause
Introduction
Rotor-Gene Q User Manual 11/2012 2-1
Introduction
2.2 General Information
2.2.1 Technical assistance
At QIAGEN we pride ourselves on the quality and availability
of our technical support. Our Technical Services Departments
are staffed by experienced scientists with extensive practical
and theoretical expertise in molecular biology and the use of
QIAGEN products. If you have any questions or experience
any difficulties regarding the Rotor-Gene Q or QIAGEN
products in general, do not hesitate to contact us.
QIAGEN customers are a major source of information
regarding advanced or specialized uses of our products. This
information is helpful to other scientists as well as to the
researchers at QIAGEN. We therefore encourage you to
contact us if you have any suggestions about product
performance or new applications and techniques.
For technical assistance and more information, call one of
the QIAGEN Technical Services Departments or local
distributors (see back cover).
For up-to-date information about the Rotor-Gene Q, visit
www.qiagen.com/RotorGeneQ.
2.2.2 Policy statement
It is the policy of QIAGEN to improve products as new
techniques and components become available. QIAGEN
reserves the right to change specifications at any time.
In an effort to produce useful and appropriate
documentation, we appreciate your comments on this user
manual. Please contact QIAGEN Technical Services.
2.2.3 Version management
This document is the Rotor-Gene Q User Manual, version 3,
for Rotor-Gene Q instruments using Rotor-Gene Q software
versions 2.1.0 or higher.
2-2 Rotor-Gene Q User Manual 11/2012
Introduction
2.3 Intended use of the Rotor-Gene Q
The Rotor-Gene Q instrument is designed to perform realtime and end-point thermal cycling using the polymerase
chain reaction (PCR) and high-resolution melting analysis
(HRM™) in molecular biology applications as well as for
other applications such as concentration measurement,
protein analysis, and enzyme kinetics.
The Rotor-Gene Q, if used in combination with QIAGEN Kits
indicated for use with the Rotor-Gene Q instrument, is
intended for the applications described in the respective
QIAGEN Kit handbooks.
If the Rotor-Gene Q instrument is used with kits other than
QIAGEN Kits, it is the user’s responsibility to validate the
performance of such product combination for any particular
application.
The Rotor-Gene Q instrument is intended for use by
professional users, such as technicians and physicians
trained in molecular biological techniques and the operation
of the Rotor-Gene Q instrument.
Rotor-Gene Q User Manual 11/2012 2-3
Introduction
This page intentionally left blank
2-4 Rotor-Gene Q User Manual 11/2012
3 General Description
Rotor chamber
The Rotor-Gene Q is an innovative instrument that enables
high-precision real-time PCR, end-point PCR, and high
resolution melt (HRM) analysis. It is highly suited for use in
gene expression analysis, genotyping, pathogen detection,
and many other areas of research.
The powerful and user-friendly software provides simplicity
for beginners as well as an open experimental platform for
advanced users.
Air vents
Lid handle
Instrument
status lights
General Description
3.1 Thermal performance
The Rotor-Gene Q uses a sophisticated heating and cooling
design to achieve optimal reaction conditions. The unique
rotary format ensures optimal thermal and optical uniformity
between samples which is critical for precise and reliable
analysis.
Samples spin continually at 400 rpm during a run.
Centrifugation prevents condensation and removes air
Rotor-Gene Q User Manual 11/2012 3-1
General Description
bubbles, but does not pellet DNA. In addition, samples do
not need to be spun down prior to a run.
Samples are heated and cooled in a low-mass–air oven.
Heating is achieved by a nickel-chrome element in the lid.
The chamber is cooled by venting the air out through the top
of the chamber while ambient air is blown up through the
base.
Heating
Cooling
Illustration of the heating and cooling system.
3-2 Rotor-Gene Q User Manual 11/2012
3.2 Optical system
With a choice of up to 6 excitation sources and 6 detection
filters combined with a short, fixed optical path, the
Rotor-Gene Q can be used for multiplex reactions, ensuring
minimum fluorescence variability between samples and
eliminating the need for calibration or compensation.
Samples are excited from the bottom of the chamber by a
light-emitting diode. Energy is transmitted through the thin
walls at the base of the tube. Emitted fluorescence passes
through emission filters on the side of the chamber and is
then collected by a photomultiplier. The fixed optical path
ensures consistent excitation for every sample, which means
that there is no need to use a passive internal reference dye
such as ROX.
General Description
Rotor-Gene Q User Manual 11/2012 3-3
Illustration of the optical system.
General Description
Red®, Alexa Fluor 568
Alexa Fluor 680
Available channels
Channel Excitation
(nm)
Detection
(nm)
Examples of
fluorophores
detected
®
Blue 365±20 460±20 Marina Blue
, Edans
Bothell Blue, Alexa
®
350, AMCA-X,
Fluor
ATTO 390
Green
470±10
510±5
FAM®, SYBR® Green I,
Fluorescein,
EvaGreen®, Alexa
Fluor 488
Yellow 530±5 557±5 JOE™, VIC®, HEX,
®
TET™, CAL Fluor
Gold 540, Yakima
®
Orange
585±5
610±5
Yellow
ROX™, CAL Fluor Red
610, Cy®3.5, Texas
Red 625±10 660±10 Cy5, Quasar® 670,
®
LightCycler
Red640,
Alexa Fluor 633
Crimson
680±5
712 high
pass
Quasar 705,
LightCycler Red705,
3-4 Rotor-Gene Q User Manual 11/2012
High
resolution
melt
(HRM)
460±20 510±5 SYBR Green I,
®
SYTO
9, LC Green®,
LC Green Plus+,
EvaGreen
Note: QIAGEN kits indicated for use with the Rotor-Gene Q
instruments are optimized with respect to certain dye
combinations. Please refer to the corresponding kit
handbooks, for example, the Rotor-Gene Multiplex
®
Handbook or the QuantiTect
Virus Handbook for more
information.
Installation Procedures
4 Installation Procedures
4.1 Site requirements
Rotor-Gene Q instruments must be located out of direct
sunlight, away from heat sources, and away from sources of
vibration and electrical interference. Refer to Appendix A for
the operating conditions (temperature and humidity). The
installation site should be free of excessive drafts, excessive
moisture, excessive dust, and not subject to large
temperature fluctuations.
Refer to Appendix A for the weight and dimensions of
Rotor-Gene Q instruments. Ensure that the workbench is dry,
clean, and has additional space for accessories. For further
information about required specifications of the workbench,
contact QIAGEN Technical Services.
Note: It is extremely important that the Rotor-Gene Q
instrument is placed on a stable surface, which is level and
vibration free. Refer to operating conditions — see
Appendix A.
The Rotor-Gene Q instrument must be placed within
approximately 1.5 m (59 in.) of a properly grounded
WARNING
WARNING
Rotor-Gene Q User Manual 11/2012 4-1
(earthed) AC power outlet.
Explosive atmosphere [W10]
The Rotor-Gene Q instrument is not designed for use in an
explosive atmosphere.
Risk of overheating [W16]
To ensure proper ventilation, maintain a minimum
clearance of 10 cm (3.94 in.) at the rear of the Rotor-Gene
Q instrument.
Slits and openings that ensure the ventilation of the
Rotor-Gene Q instrument must not be covered.
Installation Procedures
4.2 AC Power connection
Power requirements
The Rotor-Gene Q operates at:
100–240 V AC, 50/60 Hz; 560 VA (peak)
Make sure that the voltage rating of the Rotor-Gene Q is
compatible with the AC voltage available at the installation
site. Mains supply voltage fluctuations are not to exceed 10%
of nominal supply voltages.
Grounding requirements
To protect operating personnel, QIAGEN recommends that
the Rotor-Gene Q be correctly grounded (earthed). The
instrument is equipped with a 3-conductor AC power cord
that, when connected to an appropriate AC power outlet,
grounds (earths) the instrument. To preserve this protection
feature, do not operate the instrument from an AC power
outlet that has no ground (earth) connection.
Installation of AC power cord
Connect the suitable end of the AC power cord to the socket
located at the rear of the Rotor-Gene Q instrument, and the
other end to the AC power outlet.
4.3 PC requirements
The laptop computer, optionally supplied with the
Rotor-Gene Q, fulfills the requirements of the Rotor-Gene Q
4-2 Rotor-Gene Q User Manual 11/2012
software, detailed in the following table.
Loading...