Advanced User Manual
249 Pages
Preview
Page 1
March 2011
QIAxcel Advanced User Manual
Sample & Assay Technologies
Trademarks: QIAGEN®, QIAsymphony®, QIAxcel® (QIAGEN Group); Adobe®, Reader® (Adobe Systems Inc.); Excel®, Microsoft®, Windows® (Microsoft Corporation); Intel®, Celeron® (Intel Corporation). Registered names, trademarks, etc. used in this document, even when not specifically marked as such, are not to be considered unprotected by law. 1067883 03/2011 © 2011 QIAGEN, all rights reserved.
Contents
3
Contents 1
Safety Information
1-2
1.1
... 1-2 Proper use
1.2
... 1-4 Electrical safety
1.3
... 1-5 Environment
1.4
... 1-5 Chemicals
1.5
... 1-6 Waste disposal
1.6
... 1-6 Mechanical hazards
1.7
1-6 Symbols... on the QIAxcel Advanced
2
Introduction
2-2
2.1
... 2-2 About this user manual
2.2
2-2 General... information
2.3
2-3 Intended... use of the QIAxcel Advanced
2.3.1
3
Requirem ents for ... QIAxcel Advanced users 2-4
General Description
3-2
3.1
3-2 QIAxcel ... Advanced principle
3.2
3-3 External... features of the QIAxcel Advanced
3.3
... 3-4 Gel cartridges
3.4
... 3-5 Computer and software
4 4.1
Installation Procedures
4-2
... 4-2 Requirements Site ... 4-2 Pow er requirem... ents 4-2 Grounding requirem ... ents 4-3
4.1.1 4.1.2 4.1.3 4.2
... 4-3 Unpacking the QIAxcel Advanced
4.3
... 4-3 Installing the QIAxcel Advanced
4.3.1 4.3.2 4.3.3 4.3.4 4.3.5 4.3.6 4.4
5
Releasing the transport ... lock 4-4 Instrum ent setup ... 4-4 Installation of AC ... pow er cord 4-4 Installation of the ... N2 cylinder 4-5 Installation of the ... QIAxcel ScreenGel softw are 4-5 Getting started w ... ith the QIAxcel ScreenGel softw are 4-6
4-6 Packing... the QIAxcel Advanced
Operating Procedures
5-2
5.1
... 5-2 Unpacking a QIAxcel Kit
5.2
... 5-3 Setting up the QIAxcel Advanced
QIAxcel Advanced User Manual 03/2011
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4
QIAxcel Advanced User Manual Preparing the buffer ... tray 5-3 Loading the buffer ... tray 5-4 Installing a QIAxcel ... gel cartridge and sm art key 5-5 Rem oving a QIAxcel ... gel cartridge 5-6 Storing a QIAxcel ... gel cartridge 5-6
5.2.1 5.2.2 5.2.3 5.2.4 5.2.5 5.3
... 5-7 Operating the QIAxcel Advanced
5.3.1 5.3.2
Things to do before ... starting 5-7 Starting a run ... 5-8
6 6.1 6.1.1 6.1.2 6.1.3 6.1.4 6.1.5
QIAxcel ScreenGel Software
6-2
... 6-2 Concepts Modes ... 6-2 User roles ... 6-3 Environm ents ... 6-5 Profiles ... 6-5 General softw are ... usage 6-6 Editing data in tables ... 6-8 Displaying errors ... and w arnings 6-9 Displaying modifications ... 6-11 Getting help ... 6-14
6.2
... 6-14 User authentication
6.3
Process... 6-17
6.3.1 6.3.2 6.3.3
6.3.4 6.3.5 6.3.6 6.4 6.4.1
6.4.2
Running a process ... w ith advanced options 6-19 "Provide Sample ... Information" 6-25 Modifying a process ... profile 6-27 Process profile... options 6-29 Selecting general ... process options 6-31 Selecting run ... parameters 6-32 Selecting analysis ... parameters 6-35 Selecting marker ... 6-37 Selecting binary ... peak definition table 6-39 Selecting report/export ... parameters 6-40 Run parameters ... and result structure 6-41 Creating a new... process profile 6-44 View ing m ethod ... details 6-46 "Status Inform ation" ... panel 6-47
... 6-49 Analysis Handling sam ples ... and experim ents 6-50 Meaning of sample ... icons 6-52 Loading sample ... data 6-52 Selecting samples ... 6-53 Expanding and ... collapsing 6-54 Activating an ... experiment 6-54 Saving an experiment ... 6-54 Closing an experiment ... 6-55 Importing an experiment ... 6-55 Exporting an experiment ... 6-56 Importing BioCalculator ... Data 6-56 View ing sam ple ... data 6-58 Adding samples ... to view s 6-58 Removing samples ... from view s 6-60 Exporting view ... to clipboard 6-61 QIAxcel Advanced User Manual 03/2011
Contents
6.4.3
6.4.4
6.4.5
6.4.6
6.4.7
6.4.8 6.5 6.5.1
6.5.2
6.5.3
6.6 6.6.1
5
Result table ... 6-63 Gel view ... 6-66 Electropherogram ... view 6-69 Electropherogram ... overview 6-75 Electropherogram ... superposition view 6-78 Inspecting sample ... properties 6-80 Saturated signals ... 6-80 Analysis DNA ... 6-81 Analysis procedure ... DNA 6-83 Modifying an analysis ... profile 6-87 Creating a new ... analysis profile 6-91 Creating a DNA ... reference marker 6-91 Creating a new ... size marker table 6-94 Performing binary ... peak calling 6-96 Modifying a binary ... peak definition table 6-98 Creating a new ... binary peak definition table 6-99 Analysis RNA ... 6-100 Analysis procedure ... RNA 6-101 Creating a RNA ... reference marker 6-105 Manual correction ... of RNA analysis 6-108 Manually m odifying ... analysis results 6-109 Deleting a peak ... 6-109 Adding a peak ... 6-110 Removing analysis ... results 6-110 Checking alignment ... marker 6-111 Custom izing experim ... ents 6-111 Creating a new ... experiment 6-112 Modifying an... experiment 6-113 Report/Export... 6-116 Generating a... report 6-117 Exporting data ... 6-118 Modifying a report/export ... profile 6-118 Report/Export ... options 6-119 Creating a new ... report/export profile 6-128 Im port BioCalculator ... sam ple data 6-129
... 6-131 Service Calibrating a cartridge ... 6-132 Running the ... calibration w izard 6-132 Recalibrating... a cartridge 6-133 System Check... 6-134 Complete Check ... 6-134 Detector Test ... 6-135 Filter Check ... 6-136 Movement Check ... 6-137 Leakage Test ... 6-138 Maintenance ... 6-139 Purge ... 6-140 Long purge ... 6-140 Empty N2 bottle ... 6-141 Setting the instrument ... ID 6-141 Troubleshooting ... folder 6-142
... 6-143 Configuration Settings
... 6-143
QIAxcel Advanced User Manual 03/2011
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6 6.6.2 6.6.3
7
QIAxcel Advanced User Manual Profile m anagem ... ent 6-148 User m anagem ... ent 6-149
Maintenance Procedures
7-2
7.1
7-2 Cleaning... of the QIAxcel Advanced
7.2
... 7-3 Minor corrective maintenance
7.2.1 7.2.2 7.2.3 7.2.4
8
Fuse replacem ent ... and cleaning 7-3 N2 cylinder replacem ... ent 7-3 Alternate N2 supply ... 7-4 Blocked purge filter ... 7-4
Troubleshooting
8-2
8.1
... 8-2 System setup
8.2
... 8-3 Operation
8.3
... 8-3 DNA applications
8.4
... 8-6 RNA applications
9
Glossary
9-2
10
Appendices
10-2
10.1
... 10-3 Appendix A
10.2
... 10-7 Appendix B
10.3
... 10-14 Appendix C
10.4
... 10-17 Appendix D
10.5
... 10-21 Appendix E
10.6
... 10-22 Appendix F
10.7
... 10-27 Appendix G
QIAxcel Advanced User Manual 03/2011
1. Safety Information
1-2
QIAxcel Advanced User Manual
1
Safety Information Before using the QIAxcel Advanced, 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. The following types of safety information appear throughout this manual.
WARNING
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.
CAUTION
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 this one. The advice given in this manual is intended to supplement, not supersede, the normal safety requirements prevailing in the user’s country.
1.1 WARNING/ CAUTION
Proper use Risk of personal injury and material damage Improper use of the QIAxcel Advanced may cause personal injuries or damage to the instrument. The QIAxcel Advanced must only be operated by qualified personnel who have been appropriately trained. Servicing of the QIAxcel Advanced must only be performed by QIAGEN Field Service Specialists.
[W1]
Perform the maintenance as described in the Maintenance Procedures section. QIAGEN charges for repairs that are required due to incorrect maintenance.
WARNING/ CAUTION
Risk of personal injury and material damage The QIAxcel Advanced is too heavy to be lifted by one person. To avoid personal injury or damage to the instrument, do not lift the instrument alone.
[W2]
WARNING/ CAUTION
Risk of personal injury and material damage Do not attempt to move the QIAxcel Advanced during operation.
[W3]
CAUTION
Damage to the instrument
[C1]
QIAxcel Advanced User Manual 03/2011
Safety Information
Avoid spilling water or chemicals onto the QIAxcel Advanced. Damage caused by water or chemical spillage will void your warranty.
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1-4
QIAxcel Advanced User Manual
In case of emergency, switch off the QIAxcel Advanced at the power switch at the rear of the instrument and unplug the power cord from the power outlet.
CAUTION
1.2
[C2] Damage to the instrument If the "Pressure 1" status is "Low", increase system pressure before performing the "unlatch" command. Removing the cartridge and unlatching with reduced pressure may damage the instrument.
Electrical safety Disconnect the line power cord from the power outlet before servicing.
WARNING
[W4] Electrical hazard 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 QIAxcel Advanced, 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: 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.
QIAxcel Advanced User Manual 03/2011
Safety Information
1.3
1-5
Environment Operating conditions
WARNING
Explosive atmosphere The QIAxcel Advanced is not designed for use in an explosive atmosphere.
WARNING
[W6] Risk of explosion The QIAxcel Advanced is intended for use with reagents and substances supplied with QIAGEN QIAxcel Kits. Use of other reagents and substances may lead to fire or explosion.
CAUTION
Damage to the instrument Direct sunlight may bleach parts of the instrument and cause damage to plastic parts. The QIAxcel Advanced must be located out of direct sunlight.
CAUTION
[C4] Damage to the cartridge When in use, the gel cartridge should not be left out of the "Wash park" position of the solution tray for more than 15 minutes. Failure to do so will cause the capillary tips to dry out, affecting correct function of the cartridge. Dry tips will void the warranty. The capillary tips are made of glass and are very fragile. Take care not to knock the tips on any hard surfaces. Failure to do so will cause the capillary tips to break, affecting correct function of the cartridge. Broken tips will void the warranty.
1.4 WARNING
[W5]
[C3]
Chemicals [W7] Hazardous chemicals Some chemicals used with this instrument may be hazardous or may become hazardous after completion of the 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,† or 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.
* 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).
WARNING
Risk of fire When cleaning the QIAxcel Advanced with alcohol-based disinfectant, leave the QIAxcel Advanced doors open to allow flammable vapors to disperse.
QIAxcel Advanced User Manual 03/2011
[W8]
1-6
QIAxcel Advanced User Manual
1.5
Waste disposal Used labware and containers may contain hazardous chemicals. Such wastes must be collected and disposed of properly according to local safety regulations. For information about how to dispose of the QIAxcel Advanced, see Appendix A.
1.6
Mechanical hazards The cartridge door and sample door of the QIAxcel Advanced must remain closed during operation of the instrument.
WARNING
1.7
[W9] Moving parts To avoid contact with moving parts during operation of the QIAxcel Advanced, the instrument must be operated with the cartridge door and sample door closed. If the door sensors are not functioning correctly, contact QIAGEN Technical Services.
Symbols on the QIAxcel Advanced Symbol
Location Description Type plate on the back of the CE mark instrument Type plate on the back of the CSA listing mark for Canada instrument and the USA Type plate on the back of the Legal manufacturer instrument Type plate on the back of the Waste Electrical and instrument Electronic Equipment (WEEE) Type plate on the back of the FCC mark of the United instrument States Federal Communications Commission
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Safety Information
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1-7
2. Introduction
Introduction
2
2-2
Introduction Thank you for choosing the QIAxcel Advanced. We are confident it will become an integral part of your laboratory. Before using the QIAxcel Advanced, 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 QIAxcel Advanced in the following sections: Safety Information Introduction General Description Installation Procedures Operating Procedures QIAxcel ScreenGel Software Maintenance Procedures Troubleshooting Glossary Appendices
The appendices include the following: Technical data QIAxcel Advanced methods and accessories Data analysis algorithm descriptions Warranty liability clause
2.2
General information 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 sample and assay technologies and the use of QIAGEN® products. If you have any questions or experience any difficulties regarding the QIAxcel Advanced 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, please see our Technical Support Center at www. qiagen.com/goto/TechSupportCenter or call one of the QIAGEN Technical Service Departments or local distributors (see back cover or visit www.qiagen.com).
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QIAxcel Advanced User Manual
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 would appreciate your comments on this user manual. Please contact QIAGEN Technical Services.
Version management This document is the QIAxcel Advanced User Manual, version 1.0.
2.3
Intended use of the QIAxcel Advanced The QIAxcel Advanced is a multicapillary electrophoresis instrument, designed to perform fast and fully automated DNA fragment analysis or qualitative and quantitative RNA analysis. The QIAxcel Advanced instrument is intended to be used only in combination with QIAxcel Kits for applications described in the respective QIAxcel Kit handbooks. The QIAxcel Advanced instrument is intended for use by professional users, such as technicians and physicians trained in molecular biological techniques and the operation of the QIAxcel Advanced instrument.
QIAxcel Advanced User Manual 03/2011
Introduction
2.3.1
Requirements for QIAxcel Advanced users This table covers the general level of competence and training necessary for transportation, installation, use, maintenance, and servicing of the QIAxcel Advanced.
Task
Personnel
Training and experience
Transportation
No special requirements
No special requirements
Installation
Laboratory technicians or equivalent
Appropriately trained and experienced personnel familiar with use of computers and automation in general
Instrument administration
Laboratory technicians or equivalent
Appropriately trained and experienced personnel familiar with use of computers and automation in general
Routine use Laboratory technicians or (running profiles) equivalent
Appropriately trained and experienced personnel familiar with use of computers and automation in general
Analyzing data
Laboratory technicians or equivalent
Appropriately trained and experienced personnel familiar with use of computers and automation in general
Instrument maintenance
Laboratory technicians or equivalent
Appropriately trained and experienced personnel familiar with use of computers and automation in general
Servicing
QIAGEN Field Service Specialists only
QIAxcel Advanced User Manual 03/2011
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3. General Description
General Description
3
3-2
General Description The QIAxcel Advanced system performs fully automated separation of DNA and RNA fragments according to their molecular weight and can process up to 96 samples per run. The QIAxcel Advanced system consists of the QIAxcel Advanced instrument, a QIAxcel Kit (containing a QIAxcel gel cartridge and reagents), a computer, and QIAxcel ScreenGel software, which have been optimized to work together across a wide variety of applications and provide unmatched resolution, speed, and throughput for DNA fragment and RNA analysis. The QIAxcel ScreenGel software supplied with the QIAxcel Advanced provides both electropherograms and gel images of the nucleic acid separation and can be used to perform the following types of analysis: Determination of the size and concentration of DNA fragments Determination of the ratio, concentration, and quality of total RNA, and the quality of cRNA/ cDNA and fragmented RNA/DNA Note: We do not recommend determining nucleic acid concentration with the QIAxcel DNA Fast Analysis Kit.
3.1
QIAxcel Advanced principle The QIAxcel Advanced uses capillary electrophoresis to enable high resolution and sensitive separation of DNA and RNA samples. This 12-channel capillary electrophoresis instrument uses disposable, multiple-use cartridges for cost-effective analysis of up to 96 samples in as little as 25 minutes. 1.
The QIAxcel Advanced is set up with a gel cartridge, running buffer, and wash buffer, and calibrated using intensity markers.
2.
The samples for analysis (in a 96-well plate or 12-tube strips) are placed onto the sample plate holder.
3.
Required data collection settings are selected and the samples pass through the QIAxcel gel cartridge.
4.
Data is analyzed using the QIAxcel ScreenGel software.
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3.2
QIAxcel Advanced User Manual
External features of the QIAxcel Advanced
External features of the QIAxcel Advanced.
1
Sample door
8
N2 door
2
Cartridge door
9
Transport lock and buffer tray location
3
Service door
10
Sample plate holder
4
Power connection for AC connection
11
Cartridge bay
5
Tube fitting for external N2
12
Slot for smart key
supply 6
RS232 connection
13
Digital pressure display
7
N2 regulator and cylinder
14
Purge filter
Note: If using an external N2 source, the output pressure must not exceed 75 psi. The QIAxcel Advanced is equipped with an internal regulator that will regulate the pressure provided by the external N2 source to approximately 40 psi (37–45 psi), which is the instrument’s operating pressure.
QIAxcel Advanced User Manual 03/2011