Techne PrimeQ OPERATOR’S MANUAL

PrimeQ
OPERATOR’S MANUAL
Version 2.0
06/12 2
Please read all the information in this guide before using the unit.
Le rogamos lea cuidadosamente la información contenida en este folleto antes de manipular el aparato.
Veuillez lire attentivement toutes les instructions de ce document avant d’utiliser l’appareil.
This operator guide provides the basic information to get you started on the PrimeQ real-time PCR system. It covers everything from taking the instrument out of the box, installing the application software and system set-up, to setting up, running and analysing a range of real-time PCR experiments. Background information is included at the start of each chapter to help familiarize the user with the theory of operation and instrument setup.
This guide makes the assumption that the reader has a basic understanding of:
Laboratory techniques; including assay design and preparation
Microsoft Windows including terminology, common commands, file saving etc.
The guide is separated into the following main areas:
Chapter Title Content
Chapter 1 Safety and installation
information
Chapter 2 Introduction to PrimeQ
and real-time PCR
Chapter 3 Using Quansoft Introduction to Quansoft
Chapter 4 Data analysis Introduction/theory of data analysis
Instrument specifications
Warranty and certification
Site requirements; PC requirements
Installing Quansoft applications software
Filter installation
Overview of real-time PCR
Examples of fluorescent chemistries
PrimeQ system overview
PCR program setup
Plate layout setup
Analysis setup
Starting/running/stopping/editing an experiment
Quantification
Dissociation curve
Plus/minus scoring
Allelic discrimination
End-point analysis (Multi-read)
Multiplexing
Chapter 5 Technical information Maintenance and spare parts
Technical support
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Chapter 6 Troubleshooting and
Glossary
Identifying problems with the instrument and/or the PCR
Glossary of terms used in real-time PCR
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5 6
Contents
Introduction ................................................................... ......................................... ................ 13
1
1.1PrimeQ ............................................................................................................................. 15
1.1.1 PrimeQ features ...................................................................................................... 15
1.1.2 The thermal system ................................................................................................. 15
1.2Quansoft .......................................................................................................................... 15
1.2.1 Experiment Editor .................................................................................................... 15
1.2.2 Plate Layout Editor .................................................................................................. 15
1.2.3 Program Editor ........................................................................................................ 16
1.2.4 PrimeQ reports ........................................................................................................ 16
1.2.5 Data Analysis .......................................................................................................... 16
1.3Unpacking ........................................................................ ................................................ 17
1.4Warning! .......................................................................................................................... 17
1.5Uses of PrimeQ ................................................................................................................ 17
1.6Plates ......................................................... ......................................... ............................. 17
1.7Thermal seal .................................................................................................................... 18
1.8Contacting us ................................................................................................................... 18
1.9Installing the hardware ..................................................................................................... 18
1.9.1 Site requirements .................................................................................................... 19
1.9.2 Installing the instrument ........................................................................................... 19
1.9.3 Operator safety ........................................................................................................ 20
1.9.4 Installation ............................................................................................................... 20
1.10 Before switching on ..................................................................................................... 23
1.10.1PrimeQ front view .................................................................................................... 23
1.10.2Insert the block ........................................................................................................ 23
1.10.3Fuses ...................................................................................................................... 23
1.10.4Connections on the back of the unit ........................................................................ 24
1.11 Technical Specification ................................................................................................ 25
1.12 Working conditions ...................................................................................................... 26
1.13 Guarantee .................................................................................................................... 26
1.14 Installing the application software ................................................................................ 28
1.14.1Set the decimal symbol to‘.’ ..................................................................................... 28
1.14.2Loading Quansoft onto the PC ................................................................................ 29
1.14.3Registering the software .......................................................................................... 29
1.14.4Software upgrades .................................................................................................. 30
1.15 Using the LCD control panel ........................................................................................ 31
1.15.1Function keys .......................................................................................................... 31
1.15.2Power up screen ..................................................................................................... 31
1.15.3Beeper .......................................................................... ........................... ................ 31
1.15.4Before you start ....................................................................................................... 32
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Start-up procedure ................................................................................................... 32
1.15.5
1.16 Installing the filter cartridges ........................................................................................ 32
1.16.1The role of the filter cartridge ................................................................................... 32
1.16.2Filter cartridge care instructions ............................................................................... 33
1.16.3Filter cartridge installation ........................................................................................ 33
1.16.4Adding a filter cartridge ............................................................................................ 34
1.16.5Assigning filter descriptions in Quansoft .................................................................. 35
1.16.6Editing filter descriptions .......................................................................................... 35
1.16.7Removing a filter cartridge ....................................................................................... 35
1.16.8Points to remember ................................................................................................. 36
1.17 Changing the instrument name .................................................................................... 36
1.18 LED power settings ...................................................................................................... 38
1.19 Inserting a plate ........................................................................................................... 38
1.19.1Inserting a previously run plate ................................................................................ 38
1.19.2Running a plate ....................................................................................................... 38
1.20 After use ...................................................................................................................... 39
2 Getting started ....................................................................................................................... 41
2.1Introduction to PrimeQ ..................................................................................................... 43
2.1.1 Principle of a real-time PCR instrument ................................................................... 43
2.1.2 Principle of PrimeQ .................................................................................................. 43
2.1.3 Applications of PrimeQ ............................................................................................ 43
2.2Introduction to Real-time PCR ......................................................................................... 44
2.2.1 PCR ......................................................................................................................... 44
2.2.2 Qualitative vs. real-time PCR ................................................................................... 44
2.2.3 Real-time PCR on PrimeQ....................................................................................... 45
2.3Overview of fluorescent chemistries ................................................................................. 45
2.3.1 Intercalating dyes .................................................................................................... 46
2.3.2 Fluorescent labelled probes ..................................................................................... 46
2.3.3 Molecular Beacons .................................................................................................. 47
2.4System overview .............................................................................................................. 48
2.5Optics module .................................................................................................................. 48
2.5.1 The optical light path ............................................................................................... 49
2.5.2 Filter cartridges ........................................................................................................ 49
2.5.3 Adjustable LED intensity .......................................................................................... 49
2.6Thermal cycling module ................................................................................................... 49
2.7Scanning module ............................................................................................................. 50
3 Quansoft ................................................ ................................................................................ 51
3.1Introducing Quansoft ........................................................................................................ 53
3.2Software overview ............................................................................................................ 54
3.2.1 Home page .............................................................................................................. 55
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The Editors .............................................................................................................. 55
3.2.2
3.3Using Quansoft ................................................................................................................ 57
3.3.1 Home page .............................................................................................................. 57
3.3.2 Main screen ............................................................................................................. 58
3.3.3 Navigation bar ......................................................................................................... 58
3.3.4 Title bar functions .................................................................................................... 59
3.3.5 Menu bar functions .................................................................................................. 60
3.3.6 Accessing the editors .............................................................................................. 61
3.4Setting up an experiment ................................................................................................. 62
3.4.1 An Overview ............................................................................................................ 62
3.4.2 Creating a new experiment ...................................................................................... 63
3.4.3 Setting up a program ............................................................................................... 64
3.4.4 Setting up a plate layout .......................................................................................... 74
3.4.5 Defining the analysis method ................................................................................... 83
3.4.6 Saving an experiment to the library ......................................................................... 87
3.5Running an experiment .................................................................................................... 89
3.5.1 Starting the run ........................................................................................................ 89
3.5.2 Monitoring the run.................................................................................................... 92
3.5.3 Stopping or pausing a run ....................................................................................... 94
3.6LED intensity settings ...................................................................................................... 95
3.7Results Editor ................................................................................................................... 96
3.7.1 Post-run analysis main screen ................................................................................. 96
3.7.2 Viewing the results of a run ..................................................................................... 97
3.7.3 Editing a graph ........................................................................................................ 99
3.7.4 Changing the analysis parameters ........................................................................ 100
3.7.5 Log/Audit trail ........................................................................................................ 102
3.7.6 Report ................................................................................................................... 102
3.8Exporting and printing results ......................................................................................... 104
3.8.1 Exporting ............................................................................................................... 104
3.8.2 Printing .................................................................................................................. 105
4 Data analysis ....................................................................................................................... 107
4.1Introduction .................................................................................................................... 109
4.1.1 Amplification curve ................................................................................................ 109
4.1.2 Thresholds ............................................................................................................. 109
4.1.3 Fit points ................................................................................................................ 110
4.1.4 First derivative maximum ....................................................................................... 111
4.1.5 Standard curve ...................................................................................................... 111
4.1.6 Dissociation curve ................................................................................................. 111
4.2Choosing an analysis method ........................................................................................ 113
4.3Analysis method: None .................................................................................................. 115
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Viewing the results ................................................................................................ 115
4.3.1
4.3.2 PrimeQ Report ...................................................................................................... 116
4.4Passive reference dye (PRD) correction ........................................................................ 117
4.5Analysis method: Baseline correction ............................................................................ 118
4.5.1 Assay requirements ............................................................................................... 118
4.5.2 Setup .......................................................... ........................................................... 118
4.5.3 Viewing the results ................................................................................................ 120
4.5.4 PrimeQ Report ...................................................................................................... 122
4.5.5 Quick guide to baseline correction analysis ........................................................... 122
4.6Analysis method: Quantification ..................................................................................... 123
4.6.1 Assay requirements ............................................................................................... 123
4.6.2 Setup .......................................................... ........................................................... 123
4.6.3 Viewing the results ................................................................................................ 127
4.6.4 PrimeQ Report ...................................................................................................... 128
4.6.5 Using Cq values .................................................................................................... 128
4.6.6 Comparing Cq values using a standard curve ....................................................... 128
4.6.7 Comparing Cqs in relative quantification ............................................................... 132
4.6.8 Quick guide to quantification analysis .................................................................... 137
4.7Analysis method: Dissociation curve .............................................................................. 138
4.7.1 Assay requirements ............................................................................................... 139
4.7.2 Setup .......................................................... ........................................................... 139
4.7.3 Viewing the results ................................................................................................ 145
4.7.4 PrimeQ Report ...................................................................................................... 146
4.7.5 Quick guide to dissociation curve analysis ............................................................ 146
4.8Analysis method: Plus-minus scoring ............................................................................. 148
4.8.1 Assay requirements ............................................................................................... 148
4.8.2 Setup .......................................................... ........................................................... 148
4.8.3 Viewing the results ................................................................................................ 150
4.8.4 PrimeQ Report ...................................................................................................... 152
4.8.5 Quick guide to plus-minus scoring analysis ........................................................... 152
4.9Analysis method: Allelic discrimination ........................................................................... 154
4.9.1 Assay requirements ............................................................................................... 154
4.9.2 Setup .......................................................... ........................................................... 154
4.9.3 Viewing the results ................................................................................................ 156
4.9.4 PrimeQ Report ...................................................................................................... 159
4.9.5 Quick guide to allelic discrimination analysis ......................................................... 159
4.10 Analysis method: Multi-read ....................................................................................... 161
4.10.1Assay requirements ............................................................................................... 161
4.10.2Setup ....................................................................... .............................................. 161
4.10.3Viewing the results ................................................................................................ 162
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PrimeQ Report ...................................................................................................... 164
4.10.4
4.10.5Quick guide to multi-read analysis ......................................................................... 165
4.11 Multiplex assays ........................................................................................................ 166
4.11.1Multiplex setup ...................................................................................................... 166
5 Technical information ........................................................................................................... 169
5.1Operator maintenance ................................................................................................... 171
5.1.1 Cleaning PrimeQ ................................................................................................... 171
5.1.2 Fuses .................................................................................................................... 171
5.1.3 Insulation Testing .................................................................................................. 172
5.1.4 Mantenimiento ............................................................................ ........................... 172
5.1.5 Entretien utilisateur ................................................................................................ 172
5.2Block Access.................................................................................................................. 174
5.3User responsibilities ....................................................................................................... 175
5.3.1 LED ....................................................................................................................... 175
5.3.2 Filters .................................................................................................................... 175
5.3.3 Replacing the fuses ............................................................................................... 175
5.4Consumables .......................................................... ....................................................... 175
5.5Minimum computer requirements ................................................................................... 176
5.6Accessories ................................................................................................................... 176
5.7Replacement parts ......................................................................................................... 177
5.8Packing the PrimeQ instrument...................................................................................... 178
5.8.1 Remove the filter cartridges ................................................................................... 178
5.8.2 Remove the block .................................................................................................. 178
5.8.3 Packing the instrument .......................................................................................... 179
5.9Packaging ...................................................................................................................... 180
5.9.1 Returns authorization number ............................................................................... 180
5.9.2 De-contamination certificate .................................................................................. 180
6 Troubleshooting ............................................................ ....................................................... 181
6.1Troubleshooting ............................................................................................................. 183
6.2Real-time PCR Glossary ................................................................................................ 186
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1 Introduction
Safety and installation information
About this chapter
This chapter provides information on general safety aspects, definitions, advice and instructions for unpacking and installing your instrument. It also gives general information about the instrument and the control software, system requirements, basic procedures, control mechanisms and software installation.
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1.1 PrimeQ
PrimeQ - the real-time nucleic acid detection system from Techne - has been designed with the advantage of an open chemistry format that allows the end user full flexibility in the methods and research they wish to pursue. This is complimented by the user-friendly application software, Quansoft, for easy setup of experiments and rapid analysis of data.
1.1.1 PrimeQ features
The white light source and PMT detector provide an impressive excitation range of 470nm
to 650nm and a detection range of 500nm to 710nm.
Multiplex - multiple wavelengths are detectable per sample using up to four paired excitation
and emission filters housed in individual cartridge systems.
Wide dynamic range up to at least nine orders of magnitude from starting copy number and
high sensitivity detecting down to 1.0nM fluorescein and single copy templates, depending upon the assay.
1.1.2 The thermal system
96-well low-profile microplate format sealed with optical film.
Temperature-controlled optical heated lid can be set between 100°C and 115°C (or off).
Designed to minimize loss of sample and prevent sample condensation.
Ramp rates up to 2.2°C/sec.
Temperature range 4°C to 98°C.
Block uniformity of less than ±0.25°C.
1.2 Quansoft
Accompanying PrimeQ is Techne’s unique, intuitive wizard-based software, Quansoft. By employing a series of user-friendly windows accessible from the home page, Quansoft enables any real-time experiment to be created with ease.
1.2.1 Experiment Editor
By using combinations of plate layouts, thermal cycling programs and analysis parameters the Quansoft Experiment Editor allows the user easy management of experimental protocols.
1.2.2 Plate Layout Editor
Within a matter of seconds a 96-well microplate can be assigned with blanks, controls, standards, user-defined samples or unknown samples, all of which are colour-coded for ease of identification.
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1.2.3 Program Editor
Individual cycle steps, stages or temperature ramps can be added quickly and easily together with other parameters to build and display the thermal cycling program.
1.2.4 PrimeQ reports
Comprehensive, completely user-customizable reports can be generated from any of the analysis methods. All result types including graphs can be exported to Microsoft other programs currently used for publication purposes.
®
Word, PowerPoint or
1.2.5 Data Analysis
Choose the analysis types and parameters from:
None
Baseline
Quantification
Dissociation curve
Plus/minus scoring
Allelic discrimination
Multi-read (end point)
1.2.5.1 Quantification
A linearity of at least nine orders of magnitude can be achieved, allowing the user to quantify DNA down to single copy template or to achieve absolute quantification of 1.0nM fluorescein in a volume of 20µl.
1.2.5.2 Dissociation analysis
Using the easy-to-program ‘ramp’ function, PrimeQ will perform a dissociation curve that can be used to determine the temperature at which the DNA strands dissociate. It provides the user with extra confidence in genotyping experiments and in product verification analysis.
1.2.5.3 Plus/minus scoring
This analysis exploits PrimeQ’s fluorescence technology to determine with ease and accuracy the presence or absence of a PCR product in any given sample.
1.2.5.4 Allelic discrimination
Users of PrimeQ have the option of this powerful technique capable of detecting single nucleotide differences (SNPs). It can be used to discriminate between genotypes, mutations and polymorphisms within or between samples simply by comparing the fluorescence signal obtained using allele-specific, dye-labelled probes.
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1.3 Unpacking
When unpacking the unit, check that the following have been removed from the packing:
This PrimeQ operator guide
A CD containing an electronic copy of this guide and Quansoft software
The PrimeQ unit
A thermal block
Filter cartridges in box
Mains lead(s)
Guarantee card
Decontamination certificate
USB cable
Keep the original packaging in case you ever need to return the unit for service or repair. Techne accepts no responsibility for damage incurred during transportation unless the unit is correctly packed and transported in its original packaging.
The instrument weighs 25kg (55lb). Take care when lifting – this is best done by two people.
1.4 Warning!
Position the unit so that the mains on/off switch is accessible. If a safety problem should be encountered, switch off at the power socket and remove the plug from the supply.
If you are using more than one unit in close proximity to each other, there must be at least 100mm between the units to allow the cooling air to flow correctly.
1.5 Uses of PrimeQ
PrimeQ has many scientific laboratory applications. Aspects of the PCR process are claimed in U.S. Patent Nos. 5,475,610 (claims 160-163 and 167) and 6,703,236 (claims 1-6) or corresponding claims in their non-US counterparts. Use of PrimeQ in such processes is covered by licenses granted to Techne by Applied Biosystems LLC (except for use in human in vitro diagnostics).
1.6 Plates
Techne recommends only the type of low-profile plate described in this Guide using reaction volumes between 15µl and 50µl. Plates must be able to withstand the temperatures you are using without any danger of deforming to the point where they fracture. Only low-profile plates should be used. If standard height PCR plates are used, PrimeQ may be damaged and the warranty will be invalid. Likewise if you put sections of cut-up plate in the block you must balance the plate sections with empty sealed plate sections or previously used plate sections to balance the pressure on the heated lid.
During the final cool-down, a ring of condensation may form on the inside walls of the plate wells, above the liquid level but below the top of the sample block. This is not usually a cause for concern as the condensation does not form during cycling, due to the action of the heated lid.
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1.7 Thermal seal
The specification given in this Guide is based on the use of an optical heat seal. Other types of optical seal may be used; however any seal must be able to withstand the temperatures you are using without any danger of deforming to the point where it splits. The user should check the suitability of the seal to be used to ensure there is minimal sample evaporation.
Where applicable, you should use a heat sealer which seals at a temperature of approximately 170ºC.
Place the plate in the heat sealer base plate, cover it with the seal. Seal the plate for 3-4 seconds; rotate the plate 180 degrees; seal the plate again for a further 3-4 seconds.
The plate is now correctly sealed and ready for inserting into PrimeQ.
Always be certain that you use the film in the correct way. If you find that you have used the seal the wrong way up DO NOT PLACE IT IN THE PRIMEQ UNIT as it will damage the heated lid and this will invalidate the warranty.
Clean the heat sealer before you try to seal another plate.
When using the recommended brand of thermal seal, FHSFILM, and find that it has been applied the wrong way, the user is advised to send the heat sealer back to their supplier for repair.
Should the user want to attempt in-house repair, the heat sealer should be left to cool and the heating plate cleaned according to the manufacturer’s instructions.
1.8 Contacting us
If you have any questions about PrimeQ, our technical support resources can be accessed in the following ways:
Internet: Visit our websites at www.techne.com or www.techneusa.com to find out the
answers to a range of frequently asked questions.
Email: Contact PrimeQHelp@bibby-scientific.com or PrimeQHelp@techneusa.com for
technical and applications assistance. For servicing enquiries contact service@bibby-
scientific.com or service@techneusa.com in the US.
Phone: For technical support call +44 (0)1785 810433. For servicing call +44 (0)1785
810475 or +1 609 589 2560 in the US.
Bibby Scientific Ltd. Beacon Road Stone Staffordshire ST15 0SA UK Tel: +44 (0)1785 812121 Fax: +44 (0)1785 810405 E-mail: sales@bibby-scientific.com
1.9 Installing the hardware
To protect the mechanisms within the PrimeQ, the thermal cycling block is packed and transported separately from the instrument (in the main carton).
DO NOT PLUG IN THE POWER CABLE OR TRY TO SWITCH ON UNTIL YOU HAVE INSTALLED THE BLOCK.
See the instructions in sections 1.10.
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1.9.1 Site requirements
PrimeQ can operate on a laboratory bench top. The unit is 420mm wide x 518mm deep x 528mm high and requires 100mm side clearance for operation. The unit weighs 25kg (55lb).
The unit should ideally be operated in the temperature range of 18°C to 30°C out of direct sunlight and draughts. Operating humidity range should be RHa up to a maximum of 80% non-condensing.
The unit can be connected to a power supply with voltages from 100 to 230V AC.
The unit must be connected to a PC as described in section 1.14.
1.9.2 Installing the instrument
1.9.2.1 Warning
HIGH TEMPERATURES ARE DANGEROUS: they can cause serious burns to operators and ignite combustible material.
Techne has taken great care in the design of these units to protect operators from hazards, but users should pay attention to the following points:
USE CARE AND WEAR PROTECTIVE GLOVES TO PROTECT HANDS;
DO NOT put hot objects on or near combustible objects;
DO NOT operate the unit close to inflammable liquids or gases;
DO NOT place any liquid directly in your unit;
At all times USE COMMON SENSE.
1.9.2.2 Aviso
LAS TEMPERATURAS ELEVADAS SON PELIGROSAS: pueden causarle graves quemaduras y provocar fuego en materiales combustibles.
Techne ha puesto gran cuidado en el diseño de estos aparatos para proteger al usuario de cualquier peligro; aun así se deberá prestar atención a los siguientes puntos:
EXTREME LAS PRECAUCIONES Y UTILICE GUANTES PARA PROTEGERSE LAS
MANOS;
NO coloque objetos calientes encima o cerca de objetos combustibles;
NO maneje el aparato cerca de líquidos inflamables o gases;
NO introduzca ningún líquido directamente en el aparato;
UTILICE EL SENTIDO COMUN en todo momento.
1.9.2.3 Avertissement
DANGER DE TEMPERATURES ELEVEES : les opérateurs peuvent subir de graves brûlures et les matériaux combustibles risquent de prendre feu.
Techne a apporté un soin tout particulier à la conception de ces appareils de façon à assurer une protection maximale des opérateurs, mais il est recommandé aux utilisateurs de porter une attention spéciale aux points suivants:
PROCEDER AVEC SOIN ET PORTER DES GANTS POUR SE PROTEGER LES MAINS.
NE PAS poser d’objets chauds sur ou près de matériaux combustibles.
NE PAS utiliser l’appareil à proximité de liquides ou de gaz inflammables.
NE PAS verser de liquide directement dans l’appareil.
FAIRE TOUJOURS PREUVE DE BON SENS.
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1.9.3 Operator safety
All users of Techne equipment must have available the relevant literature needed to ensure their safety.
It is important that only suitably trained personnel operate this equipment in accordance with the instructions contained in this Guide and with general safety standards and procedures. If the equipment is used in a manner not specified by
Techne, the protection provided by the equipment to the user may be impaired.
All Techne units have been designed to conform to international safety requirements and are fitted with an over-temperature cut-out.
If a safety problem should be encountered, switch off at the power switch and remove the plug from the supply.
1.9.3.1 Seguridad del usuario
Todos los usuarios de equipos Techne deben disponer de la información necesaria para asegurar su seguridad.
De acuerdo con las instrucciones contenidas en este manual y con las normas y procedimientos generales de seguridad, es muy importante que sólo personal debidamente capacitado opere estos aparatos. De no ser así, la protección que el equipo le proporciona al usuario puede verse reducida.
Todos los equipos Techne han sido diseñados para cumplir con los requisitos internacionales de seguridad y traen incorporados un sistema de desconexión en caso de sobretemperatura. En algunos modelos el sistema de desconexión es variable, lo que le permite elegir la temperatura según sus necesidades. En otros, el sistema de desconexión viene ya ajustado para evitar daños en el equipo.
En caso de que surgiera un problema de seguridad, desconecte el equipo de la red.
1.9.3.2 Sécurité de l’opérateur
Tous les utilisateurs de produits Techne doivent avoir pris connaissance des manuels et instructions nécessaires à la garantie de leur sécurité.
Important : cet appareil doit impérativement être manipulé par un personnel qualifié et utilisé selon les instructions données dans ce document, en accord avec les normes et procédures de sécurité générales. Dans le cas où cet appareil ne serait pas utilisé selon les consignes précisées par Techne, la protection pour l’utilisateur ne serait alors plus garantie.
Tous les appareils Techne sont conçus pour répondre aux normes de sécurité internationales et sont dotés d’un coupe-circuit en cas d’excès de température. Sur certains modèles, ce coupe­circuit est réglable pour s’adapter à l’application désirée. Sur d’autres modèles, il est pré-réglée en usine pour assurer la protection de l’appareil.
Dans le cas d’un problème de sécurité, coupez l’alimentation électrique au niveau de la prise murale et enlevez la prise connectée à l’appareil.
1.9.4 Installation
1. All Techne units are supplied with a mains cable. This is a plug-in lead type cable.
2. Before connecting the power supply, check the voltage against the rating plate. Connect the mains cable to a suitable supply according to the table below.
Note that the unit must be earthed to ensure proper electrical safety.
100V to 230V
Live Black
Neutral White
Earth Green
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The fused plug supplied with the mains cable is fitted with a 10 amp fuse to protect the instrument and the operator.
The rating plate is on the rear of the unit.
Note: the unit can work in the range 100V to 230V.
3. Plug the mains cable into the socket on the rear of the unit.
4. Place the unit on a suitable bench or flat workspace, or in a fume cupboard if required, ensuring that the air inlet vents on the underside are free from obstruction.
5. Symbols on or near the power switch of the unit have the following meanings:
O: power switch Off
I: power switch On
1.9.4.1 Instalación
1. Todos los aparatos Techne se suministran con un cable de alimentación. Puede ser fijo o independiente del aparato.
2. Antes de conectarlo, compruebe que el voltaje corresponde al de la placa indicadora. Conecte el cable de alimentación a un enchufe adecuado según la tabla expuesta a continuación.
El equipo debe estar conectado a tierra para garantizar la seguridad eléctrica.
100V - 230V
Linea Negro
Neutro Blanco
Tierra Verde
El fusible (10A) una vez instalado protege tanto al equipo como al usuario.
Los equipos funcionan a 100V y 230V.
La placa indicadora está situada en la parte posterior del equipo.
3. Conecte el cable a la toma de tensión en la parte posterior del equipo.
4. Sitúe el aparato en un lugar apropiado tal como una superficie de trabajo plana, o si fuera necesario incluso en una campana con extractor de humos, asegurándose de que las entradas de aire en la parte inferior no queden obstruidas.
5. Los símbolos que se encuentran en o cerca del interruptor de alimentación tienen los siguientes significados:
O: Interruptor principal apagado
I: Interruptor principal encendido
1.9.4.2 Installation
1. Tous les appareils Techne sont livrés avec un câble d’alimentation qui peut être intégré à l’appareil ou à raccorder.
2. Avant de brancher l’appareil, vérifiez la tension requise indiquée sur la plaque d’identification. Raccordez le câble électrique à la prise appropriée en vous reportant au tableau ci-dessous.
Il est important que l’appareil soit relié à la terre pour assurer la protection électrique requise.
100V - 230 V
Phase Noir
Neutre Blanc
Terre Vert
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Le fusible (10A) à l’intérieur de l’appareil est destiné à assurer la protection de l’appareil et de l’opérateur.
Les appareils fonctionner sur 100V et 230V
La plaque d’identification se trouve à l’arrière de l’appareil.
3. Raccordez le câble d’alimentation à la prise située à l’arrière de l’appareil.
4. Placez l’appareil sur un plan de travail ou surface plane, ou le cas échéant, dans une hotte d’aspiration, en s’assurant que les trous d’aération situés sous l’appareil ne sont pas obstrués.
5. Les symboles situés sur ou à côté de l’interrupteur de l’appareil ont la signification suivante:
O: Arrêt
l: Marche
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1.10 Before switching on
1.10.1 PrimeQ front view
1.10.2 Insert the block
Push firmly on the front of the drawer to open it and insert the block.
Slide the quick release handles in the
direction shown on the top to the unlock position.
Lift the block assembly into the unit. Never
lift or carry the block by one side, always use both sides (or support the block from underneath).
Ensure the block is fully engaged by
pushing all four corners of the block down into the carrier.
Slide the quick release handles in the opposite direction to lock the block assembly in
position.
Push the drawer back into the unit.
1.10.3 Fuses
Ensure that the correct fuses are fitted for your voltage supply.
Fuses should only be changed by a competent person with training if necessary.
For voltages from 110 V to 130 V, use T10A fuses only.
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1.10.4 Connections on the back of the unit
There are two cable connections: the mains power switch and the USB socket located on the rear of the PrimeQ.
A. USB computer connection. Plug the USB lead into the
socket and connect the other end to your computer. The lead must be less than 2 meters in length (the lead supplied with the instrument is correct).
B. Power switch.
C. Fuses
D. Power connection. Ensure that the switch is in the off, O,
position. Plug the mains cable into the power socket.
When PrimeQ is sited in the correct position and all the connections have been made, turn the switch to the on (I) position on the PrimeQ.
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1.11 Technical Specification
Thermal cycler
Block Format: 96 x 0.2ml well low-profile micro tube plate
Block Specification: 8 x Peltier block employing quad circuit technology
Block Uniformity at 50ºC: < ± 0.25ºC
Maximum Heating Ramp Rate: 2.2ºC/sec
Temperature Range: 4ºC to 98ºC
Sample Volume: 15µl to 50µl
Heated Lid: Adjustable between 100ºC and 115ºC or off
Optical detection system
Excitation Source: White LED
Detector: Photon counting photo multiplier tube (PMT)
Multiplex Dye Detection: Up to four dyes per reaction tube
to enhance performance
User Selected Filters: Maximum of four paired excitation/emission filter
Fluorescence Excitation Range: 470nm to 650nm
Fluorescence Detection Range: 500nm to 710nm
Dynamic Range: At least nine orders of magnitude from starting
Sensitivity: 1.0nM fluorescein in a 20µl sample
Dimensions
Weight: 25kg
Size and Footprint: 420mm x 528mm x 518mm (W x H x D)
Power Supply: 100V to 230V, 50/60Hz
Input Power: 715W
IP code: IP30
cartridges suitable for use with currently available dyes
copy number.
Single initial template copy detection*
* Assay dependent
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Computer requirements (Not Supplied)
The following are the recommended minimum PC specifications required for running PrimeQ:
CPU: Core 2 Duo or equivalent
Memory: 2Gb DDR RAM as a minimum
Storage: 40Gb DMA hard drive
Display: 1024 x 768 resolution minimum, 17 inch digital
monitor recommended.
Drive: DVD/CD-RW drive
®
Operating System: Microsoft
Windows
Connections: USB
Windows® XP Professional SP3 or later,
®
Vista, Windows® 7
Also useful:
Sound: Built-in sound, video and LAN facilities
Internet: Ethernet connection
Software: Internet Explorer, Microsoft® Office
PC must be operated in ‘always on’ mode, no hibernation or sleep mode allowed.
1.12 Working conditions
PrimeQ is designed to work safely under the following conditions:
Ambient temperature range: 5°C to 40°C
Humidity: Up to 80% relative humidity, non-condensing.
Notes:
The control specifications are quoted at an ambient temperature of 20°C.
The specification may deteriorate outside an ambient temperature range of 18°C to 30°C.
Warning:
The unit may be damaged if used in an ambient temperature above 40°C.
1.13 Guarantee
Please read this important GUARANTEE information.
Notwithstanding the description and specification(s) of the units contained in the Operator Guide, Techne hereby reserves the right to make such changes as it sees fit to the units or to any component of the units.
This Guide has been prepared solely for the convenience of Techne customers and nothing in this Manual shall be taken as a warranty, condition or representation concerning the description, merchantability, fitness for purpose or otherwise of the units or components.
The PrimeQ unit is guaranteed for a period of 2 years from the date of purchase.
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Within these periods we undertake to supply replacements free of charge for parts which may on examination prove to be defective, provided that the defect is not the result of misuse, accident or negligence.
On all correspondence, please quote the Serial Number in full and/or the Sales Order Number.
Any instrument requiring service under this guarantee should be taken to the supplier through whom it was purchased, or, in the case of difficulty, it should be carefully packed in its original packing and consigned, carriage paid, to us. Techne takes no responsibility for returned goods damaged in transit.
Returned goods will not be processed without a Returns Authorization Number.
Call the Service Department on +44 (0)1785 810475 or +1 609 589 2560 in the US for a Returns Authorization Number.
Please write the Returns Authorization Number on the outside of any packing and ensure that a copy of a Decontamination Certificate is visible.
Please register online or complete and return the Registration Card to the address on the card. Returning the card or registering will help us to contact you with new information about Techne products and product up-dates thus improving our service to you.
WARRANTY
Void
IF BROKEN
The Guarantee will be rendered invalid if any of the small labels shown opposite are broken or missing when the unit arrives at the supplier’s under any claim for warranty repairs.
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1.14 Installing the application software
Always set your PC or laptop from which you are running PrimeQ so that standby is switched OFF. This can easily be achieved through the control panel or:
Right click on the desktop and select Properties.
Select the Screen Saver tab and click on the Power button.
Activate the power scheme entitled Always On from the drop down menu and click Apply.
You may need to restart your computer.
1.14.1 Set the decimal symbol to‘.’
Before installing this software please ensure that the decimal symbol on the computer is set to ‘.’. If it is not then follow these instructions:
Click on the Start button:
Go to Control Panel:
Select Regional and Language Options:
Ensure the decimal in the number section
is set to a ‘.’
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1.14.2 Loading Quansoft onto the PC
To install the Quansoft software onto the PC, follow these five basic steps:
1. Turn on the PC and log on as a user having administrator rights (to be able to install software).
2. Insert the Quansoft CD into the CD-ROM drive and wait for the auto-run function to automatically start the setup (should auto-run fail to start, then the CD must be opened from Explorer and the setup program (Setup.exe) initiated manually by double-clicking).
3. Click on the Install button and wait for the installation to complete. Quansoft files will automatically be installed to C:\Programs\Techne\Quansoft. If you wish to change the destination click the Browse button and choose accordingly. You will also be given the option of creating a shortcut icon from the desktop for easy access.
4. Remove the disk from the drive.
5. Restart the computer, log on and then launch Quansoft from the Programs menu or via the shortcut icon on the desktop.
The software can be easily uninstalled from the computer: simply access the PC Control panel, click on Add/remove programs and then select Quansoft.
If upgrading from one release of Quansoft to another, then the previous release should first be uninstalled as described.
Note: the Release number should not be confused with the version numbers which relate to each of the Quansoft editors.
Techne
Techne
Techne
Bibby Scientific
Bibby Scientific
Bibby Scientific
1.14.3 Registering the software
A registration welcome screen will appear when launching Quansoft for the first time. By registering your software with Techne, you will be eligible for technical support and software updates posted on the Techne website. However, the software can still be used for a 28-day trial period without registration – simply click Register Later and the prompt screen will appear next time the software is opened. On pressing Register Now, a form will be displayed requiring the following fields to be completed: name, organization, unlock code and the instrument serial number, should one be connected.
The unlock code can be obtained in one of three ways:
1. Internet: Clicking on www.techneusa.com/PrimeQregister in the registration panel, you will be taken directly to the PrimeQ registration page.
a. Enter your details as listed on the screen.
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b. An unlock code will be e-mailed to you within one working day.
Ensure you are online before attempting to do this.
2. E-mail: PrimeQhelp@bibby-scientific.com or PrimeQHelp@techneusa.com.
Send an email complete with details of your name, company or institution, mailing address, e­mail address, telephone number, instrument and block serial number (if you have a PrimeQ) together with the unique registration number.
Exit the registration procedure by pressing ’cancel’ and return when the unlock code has been returned by email.
3. Phone: Provide us with your details and registration number by telephoning: +44 (0)1785
810433.
1.14.4 Software upgrades
Development of the PrimeQ control and analysis software is an ongoing process. To check your version number, click ‘Help/About Quansoft’ and a splash screen displaying the current details will appear.
Check the Techne websites at www.techne.com or www.techneusa.com for the most current software updates. These updates can be downloaded free of charge for registered users.
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