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Qiagen PyroMark Q96 ID User Manual

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Version 5
January 2016
®
PyroMark
Q96 ID User Manual
Sample & Assay Technologies

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  • Page 1 Version 5 January 2016 ® PyroMark Q96 ID User Manual Sample & Assay Technologies...

  • Page 3: Table Of Contents

    Contents Contents Safety Information Proper use Electrical safety Chemical hazards Mechanical hazards Heat hazards Symbols used on the PyroMark Q96 ID Introduction About this user manual General information 2.2.1 Technical assistance 2.2.2 Policy statement 2.2.3 Version management Intended use of the PyroMark Q96 ID 2.3.1...
  • Page 4 Computer and software Installation Procedures Instrument delivery and installation Requirements Moving the PyroMark Q96 ID Reinstalling the PyroMark Q96 ID: Stand alone Reinstalling the PyroMark Q96 ID: Network installation Connecting mains power Setting up network properties 4.7.1 Used IP addresses for instrument and computer...
  • Page 5 Operator’s and office computers 6.1.2 Printer 6.1.3 Data storage device Installing PyroMark Q96 Software Uninstalling PyroMark Q96 Software Upgr ading from PyroMark Q96 ID Software 6.4.1 Installing the PyroMark Data Converter Tool 6.4.2 Using the PyroMark Data Converter Tool Data backup Installation testing...
  • Page 6 Troubleshooting Unexpected results Sequence errors Peak errors Instrument-related errors Vacuum workstation-related errors Glossary Appendix A Technical data Environmental conditions Waste Electrical and Electronic Equipment (WEEE) FCC declaration Appendix B Assay design and validation Contents-4 PyroMark Q96 ID User Manual 01/2016...
  • Page 7 Contents Appendix C Liability clause Appendix D Informations de sécurité Appendix E Sicherheitshinweise Index Index-1 PyroMark Q96 ID User Manual 01/2016 Contents-5...
  • Page 8 Contents This page intentionally left blank Contents-6 PyroMark Q96 ID User Manual 01/2016...

  • Page 9: Safety Information

    Safety Information Safety Information Before using the PyroMark Q96 ID, 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.

  • Page 10: Electrical Safety

    Risk of personal injury and material damage [W2] CAUTION The PyroMark Q96 ID is too heavy to be lifted by one person. To avoid personal injury or damage to the instrument, do not lift the instrument alone. Perform the maintenance as described in Section 7. QIAGEN charges for repairs that are required due to incorrect maintenance.

  • Page 11: Chemical Hazards

    ACGIH: American Conference of Government Industrial Hygienists (United States of America). ‡ COSHH: Control of Substances Hazardous to Health (United Kingdom). Mechanical hazards The lid of the PyroMark Q96 ID must remain closed during operation of the instrument. WARNING Moving parts...

  • Page 12: Heat Hazards

    10 cm (4 in.) at the sides and rear of the PyroMark Q96 ID. Slits and openings that ensure the ventilation of the PyroMark Q96 ID must not be covered. Symbols used on the PyroMark Q96 ID Symbol...
  • Page 13 Type plate on CSA listing mark for the back of Canada and the USA the instrument PyroMark Q96 ID User Manual 01/2016...
  • Page 14 Safety Information This page intentionally left blank PyroMark Q96 ID User Manual 01/2016...

  • Page 15: Introduction

    Thank you for choosing the PyroMark Q96 ID. We are confident it will become an integral part of your laboratory. Before using the PyroMark Q96 ID, it is essential that you read this user manual carefully and pay particular attention to the safety information.

  • Page 16: General Information

    If you have any questions or experience any difficulties regarding the PyroMark Q96 ID 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.

  • Page 17: Intended Use Of The Pyromark Q96 Id

    The PyroMark Q96 ID System is intended for use by professional users, such as technicians and physicians trained in molecular biological techniques and the operation of the PyroMark Q96 ID System.

  • Page 18: Requirements For Pyromark Q96 Id Users

    Introduction 2.3.1 Requirements for PyroMark Q96 ID users The table below covers the general level of competence and training necessary for transportation, installation, use, maintenance, and servicing of the PyroMark Q96 ID. Task Personnel Training and experience Delivery No special requirements...

  • Page 19: General Description

    PyroMark Q96 Vacuum Workstation: Vacuum  Workstation only PyroMark Q96 ID: Instrument, software, and installation  PyroMark Q96 ID System: All of the above, plus any  PyroMark kits Pyrosequencing principle Pyrosequencing uses sequencing by synthesis for accurate and quantitative analysis of DNA sequences.
  • Page 20 8. Nucleotides are added one at a time. Note: Deoxyadenosine alfa-thio triphosphate (dATPαS) is used instead of natural deoxyadenosine triphosphate (dATP) since it is used efficiently by the DNA polymerase, but not recognized by the luciferase. PyroMark Q96 ID User Manual 01/2016...

  • Page 21: Pyromark Q96 Id Principle

    Pyrogram. PyroMark Q96 ID principle The PyroMark Q96 ID performs DNA sequencing using Pyrosequencing technology. 1. After setting up the run in the PyroMark Q96 Software, the PyroMark Q96 Plate Low containing DNA samples is...
  • Page 22 14. A period of 65 seconds (cycle time) is allowed between the addition of each nucleotide to ensure that all enzymatic reactions are completed. 15. The programmed sequence of nucleotide addition continues until the end of the program has been reached. PyroMark Q96 ID User Manual 01/2016...

  • Page 23: External Features Of The Pyromark Q96 Id

    External features of the PyroMark Q96 ID The PyroMark Q96 ID. Instrument lid Standby switch Indicator LED Note: The PyroMark Q96 ID is delivered with a laptop to serve as the operator’s computer (see Section 3.8). PyroMark Q96 ID User Manual 01/2016...

  • Page 24: Internal Features Of The Pyromark Q96 Id

    General Description Internal features of the PyroMark Q96 ID Internal view of the PyroMark Q96 ID. Dispensing unit Process chamber Process chamber The process chamber includes a Peltier heating block that maintains the correct temperature of the PyroMark Q96 Plate Low and its contents.

  • Page 25: Controls And Indicators

     grounded mains power outlet Keyboard: Keyboard interface for service purposes. This  should only be used by a certified QIAGEN Field Service Specialist Mouse: Mouse interface for service purposes. This should  only be used by a certified QIAGEN Field Service...

  • Page 26: Computer And Software

    QIAGEN Field Service Specialist Computer and software The PyroMark Q96 ID is shipped with a laptop that serves as the operator’s computer. The computer used for data analysis should have the following as minimum specifications: Microsoft ®...

  • Page 27: Installation Procedures

    Installation Procedures Installation Procedures Instrument delivery and installation The unpacking and installation of the PyroMark Q96 ID is carried out by a certified QIAGEN Field Service Specialist. A member of your group who is familiar with laboratory and computer equipment should be present during the installation.
  • Page 28  230 V AC, 50 Hz, 0.6 A  Ensure that the voltage rating of the PyroMark Q96 ID and PyroMark Q96 Vacuum Workstation is compatible with the AC voltage available at the installation site. Mains supply voltage fluctuations are not to exceed 10% of nominal supply voltages.

  • Page 29: Moving The Pyromark Q96 Id

    WARNING Risk of personal injury and material damage [W2] The PyroMark Q96 ID is too heavy to be lifted by one person. To avoid personal injury or damage to the instrument, do not lift the instrument alone. If you need to transport the PyroMark Q96 ID, package the instrument as follows: 1.

  • Page 30: Reinstalling The Pyromark Q96 Id: Stand Alone

    Network installation The operator’s computer is configured with a built-in Ethernet port to connect to the PyroMark Q96 ID and an external USB adaptor to connect to a Local Area Network (LAN). The main advantages of connecting the PyroMark Q96 ID to a...

  • Page 31: Connecting Mains Power

    (press the lower part of the switch). 2. Connect the mains cable to the inlet on the rear of the PyroMark Q96 ID. Connect the mains cable to a grounded mains power outlet. The PyroMark Q96 ID will automatically adjust to the correct voltage.
  • Page 32 Installation Procedures This page intentionally left blank PyroMark Q96 ID User Manual 01/2016...

  • Page 33: Operating Procedures

    Operating Procedures Operating Procedures This section describes how to operate the PyroMark Q96 ID. Before proceeding, it is recommended that you familiarize yourself with the features of the PyroMark Q96 ID by referring to Sections 3.4 and 3.5. Required items The following items are needed to perform a run: Operator’s computer loaded with PyroMark Q96...

  • Page 34: Starting The Pyromark Q96 Software

    4. Click in the toolbar. Note: Before running your samples, validate your assay using a reference DNA sample. Note: When using QIAGEN kits, use the settings stated in the kit handbooks. PyroMark Q96 ID User Manual 01/2016...

  • Page 35: Setting Up A Run

    A well is colored according to the assay loaded into the well. Alternatively, right-click a well or selection of wells and select “Load Assay”. PyroMark Q96 ID User Manual 01/2016...

  • Page 36: Managing Instrument Parameters

    QIAGEN are used. Important: In order to access instrument parameter settings online, you must register your instrument with a valid serial number (see back of instrument) at www.qiagen.com. Managing instrument parameters in PyroMark Q96 Software: 1. Select “Instrument Parameters” from the “Tools” menu to open the instrument parameters dialog box.
  • Page 37 “Instrument Parameters” dialog box. 6. Save the new instrument parameter file. 7. When creating a new Run Setup, select this new instrument parameter file from the list of instrument parameter files. PyroMark Q96 ID User Manual 01/2016...

  • Page 38: Sample Preparation

    96-well PCR plate and adhesive foil or strip caps  Biotinylated PCR template  Streptavidin Sepharose ® HP (GE Healthcare, cat. no.  17-5113-01) Sequencing primer  High-purity water (Milli-Q 18.2 MΩ x cm or equivalent)  Ethanol (70%)  PyroMark Q96 ID User Manual 01/2016...

  • Page 39: Assembly Of The Pyromark Q96 Vacuum Workstation

    Note: The liquid waste container must be able to withstand absolute vacuum. 5.4.2 DNA amplification Amplify the DNA to be analyzed by PCR using one of the primers biotinylated. See Appendix B for details on assay design and validation. PyroMark Q96 ID User Manual 01/2016...

  • Page 40: Immobilization Of The Pcr Product To Beads

    PCR plate according to the plate setup (see Section 5.3.1). 4. Add 5–20 µl of a well-optimized, biotinylated PCR product to each well used. Note: The total volume per well should be 80 µl. PyroMark Q96 ID User Manual 01/2016...

  • Page 41: Strand Separation

    * 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). PyroMark Q96 ID User Manual 01/2016...
  • Page 42 A suggested setup is shown on the next page. Refill the troughs to these levels whenever necessary. The total volume that can be added to each trough is 180 ml. There is a mark on each trough indicating 90 ml. 5-10 PyroMark Q96 ID User Manual 01/2016...
  • Page 43 Parking Position. 8. Add 40 µl of 0.4 µM sequencing primer in Annealing Buffer to each well of the PyroMark Q96 Plate Low that is to be used. PyroMark Q96 ID User Manual 01/2016 5-11...
  • Page 44 15. Transfer the tool to the trough containing Wash Buffer (trough 3). Flush the filter probes for 10 s. 16. Raise the tool to beyond 90° vertical for 5 s, to drain liquid from the filter probes (see image). 5-12 PyroMark Q96 ID User Manual 01/2016...
  • Page 45 Note: The filter probes should be left to air dry when not in use, i.e., empty the trough and ensure there is no water in the Parking Position when the tool is left there. PyroMark Q96 ID User Manual 01/2016 5-13...

  • Page 46: Annealing Of Sequencing Primer To Samples

    Note: It is important that the reagents are pipetted into the correct reagent cartridge compartments. 4. Place the reagent cartridge with the label facing towards you. Pipet the recommended volume of substrate solution into the S compartment of the reagent cartridge. 5-14 PyroMark Q96 ID User Manual 01/2016...

  • Page 47: Run

    2. Open the process chamber lid. 3. Open the plate-holding frame. 4. Place the PyroMark Q96 Plate Low on the heating block. 5. Close the plate-holding frame and the process chamber. PyroMark Q96 ID User Manual 01/2016 5-15...

  • Page 48: Loading The Reagent Cartridge

    2. Insert the filled reagent cartridge into position. It will only fit in one orientation; the label should be facing the user. 3. Close the dispensing unit cover. Ensure the latch snaps into its locked position. 4. Close the instrument lid. 5-16 PyroMark Q96 ID User Manual 01/2016...

  • Page 49: Starting The Run

    (A, C, G, or T) dispensed to the well. A legend indicates which reagent corresponds to each color. Mixer frequency, temperature of the process chamber, and pressure in the dispensing unit are displayed in real-time in the “Instrument Status Monitor”. PyroMark Q96 ID User Manual 01/2016 5-17...

  • Page 50: Post-Run Procedures

    PyroMark Q96 Software has four analysis modes: AQ, SNP, CpG, and SQA. To toggle between the modes, select “AQ”, “SNP”, “CpG”, or “SQA” in the toolbar. Note: How the analysis is performed can be modified using the “Analysis Setup” tab. 5-18 PyroMark Q96 ID User Manual 01/2016...

  • Page 51: Viewing The Analysis Results

    SQA assay) are displayed in the “Pyrogram” pane. The well information (including analysis warnings) is listed in the “Well Information” area. If several wells are selected in the plate overview, information for the well with the orange selection frame is shown. PyroMark Q96 ID User Manual 01/2016 5-19...
  • Page 52 CpG assays Select to show the mean methylation percentage of all CpG sites in the well. 5-20 PyroMark Q96 ID User Manual 01/2016...
  • Page 53 (AQ, SNP, and CpG assays only). AQ, SNP, and CpG analysis results Allele frequencies (AQ analysis), genotypes (SNP analysis), and methylation percentages (CpG analysis) are displayed at PyroMark Q96 ID User Manual 01/2016 5-21...

  • Page 54: Analysis Reports

    PyroMark Q96 Software User Guide (press the “F1” key when in the PyroMark Q96 Software). Note: To view reports generated in PDF format, a PDF ® ® reader must be installed on the computer. Adobe Reader can be downloaded at www.adobe.com. 5-22 PyroMark Q96 ID User Manual 01/2016...

  • Page 55: Finishing Work

    Note: Be sure to observe all federal, state, and local environmental regulations for the disposal of laboratory waste. 5.7.2 Leaving the instrument on To ensure stable operating conditions, the PyroMark Q96 ID should remain on between runs. PyroMark Q96 ID User Manual 01/2016 5-23...

  • Page 56: Exiting Pyromark Q96 Software

    * 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). 5-24 PyroMark Q96 ID User Manual 01/2016...
  • Page 57 4. Empty the waste container. Note: The cap can be removed without disconnecting the tubing. 5. If the vacuum workstation must be cleaned (e.g., dust or spillage), follow the instructions in Section 7.7. PyroMark Q96 ID User Manual 01/2016 5-25...
  • Page 58 Operating Procedures This page intentionally left blank 5-26 PyroMark Q96 ID User Manual 01/2016...

  • Page 59: Pyromark Q96 Software

    For more information, contact your system administrator. 1. Close any programs running on the computer. 2. Insert the PyroMark Q96 ID Application Software 2.5 CD into the CD drive. 3. In the CD menu, click “Install PyroMark Q96 Software”.

  • Page 60: Uninstalling Pyromark Q96 Software

    4. Click “Remove” (XP) or “Uninstall” (7). Upgrading from PyroMark Q96 ID Software Previous versions of PyroMark Q96 ID Software stored Pyrosequencing runs in an Oracle database. Users upgrading to the new PyroMark Q96 Software must first use the PyroMark Data Converter Tool to convert this run database into a format that is compatible with the new software.

  • Page 61: Installing The Pyromark Data Converter Tool

    Using the PyroMark Data Converter Tool The PyroMark Data Converter tool converts Pyrosequencing runs set up and completed with the old PyroMark Q96 ID Software into a file format that is compatible with the new PyroMark Q96 Software. This tool copies the run data, converts the data to the new format, and exports the converted data to the location specified by the user.
  • Page 62 Note: The database connection can be disconnected by clicking “Disconnect”. 3. Select Pyrosequencing data for conversion by checking the box next to individual runs or folders, or using the “Select all data” or “Deselect all data” buttons. PyroMark Q96 ID User Manual 01/2016...

  • Page 63: Data Backup

    Stand-alone systems and LAN systems should be tested after installation using the test method below. This test will check the main functions of the hardware and software. If the test functions correctly, the PyroMark Q96 ID can be used for running samples. PyroMark Q96 ID User Manual 01/2016...
  • Page 64 2. Press “Ctrl” + “Alt” + “Delete” and log on as the Administrator. No password is required. The Windows desktop will appear. 3. Start the PyroMark Q96 ID using the standby switch on the front panel. All lights on the front panel will illuminate. The instrument software is loaded onto the PyroMark Q96 ID.
  • Page 65 Note: If the instrument name does not appear, click “Find Instrument” to have the software detect the connected PyroMark Q96 ID Instrument. Upon detecting the instrument, the software will enter the instrument serial number and URL. Assign a name to the instrument and click “Save”.
  • Page 66 Pyrogram should be about 4000 for all wells. 25. Click “Stop” in the run progress bar. Click “Yes” in the dialog box that appears. The run will be stopped immediately and the data will be saved. PyroMark Q96 ID User Manual 01/2016...

  • Page 67: Maintenance

    Important: Disconnect the instrument from mains power before cleaning. Servicing The PyroMark Q96 ID is supplied with a warranty that lasts for 1 year, beginning on the date of shipment. The warranty includes all repairs due to mechanical breakdown. Application development, software upgrades, accessories, and disposable items are not included in the warranty.

  • Page 68: Shutdown Procedure

    ID and check that the Power indicator lamp is turned off. Routine maintenance of the PyroMark Q96 ID The PyroMark Q96 ID should be checked and cleaned regularly to remove dust and spillages. Important points before starting: Avoid harsh cleaners and chemicals, and getting ...

  • Page 69: Cleaning The Lens Array

    1. Shut down the instrument, see Section 7.1. 2. Open the instrument lid. 3. Clean each well hole in turn with a cotton swab moistened with 70% ethanol. 4. Allow the heating block to dry before closing the instrument lid. PyroMark Q96 ID User Manual 01/2016...

  • Page 70: Changing The Fuses

    Maintenance Changing the fuses The fuses of the PyroMark Q96 ID should be checked if the instrument power indicator is not illuminated, even though the power cable is connected to the mains power outlet and the standby switch is on.

  • Page 71: Cleaning And Testing The Reagent Cartridge

    (wear powder-free gloves). A jet of water should come straight out of the tip of the needle. 5. If a needle is blocked, wet the needle tip with a drop of high-purity water. PyroMark Q96 ID User Manual 01/2016...

  • Page 72: Clogged Cartridge Cleaning Procedure

    3. Check that the jet of water is straight in the direction of the needle. If not, fill with water again and press firmly. PyroMark Q96 ID User Manual 01/2016...
  • Page 73 5. After a cartridge has passed the function test, allow it to dry completely at ambient temperature (15–25°C) before using. Cleaning the PyroMark Q96 Cartridge. Filling the compartments with high-purity water. Placing pressure on compartments. Cartridge immersed in a beaker with high-purity water. PyroMark Q96 ID User Manual 01/2016...

  • Page 74: Cleaning The Pyromark Q96 Vacuum Workstation

    The following items are required: Powder-free gloves  High-purity water (Milli-Q 18.2 MΩ x cm or equivalent)  A mild detergent (if necessary)  Clean, lint-free cloths  PyroMark Q96 ID User Manual 01/2016...

  • Page 75: Testing And Replacing The Filter Probes

    10 s. 7. If the wells are not empty after 10 s, repeat the procedure from step 1. If the function test fails twice, replace the filter probes (see below). PyroMark Q96 ID User Manual 01/2016...

  • Page 76: Replacing Filter Probes

    If the tubing is broken or distorted, replace it. Be sure to observe all federal, state, and local environmental regulations for the disposal of laboratory waste. The following items are required: New tubing (contact QIAGEN Technical Services)  Beaker  7-10 PyroMark Q96 ID User Manual 01/2016...
  • Page 77 Use the old tubing as a template for the size of the new tubing pieces. Ensure that the tubing is connected to the pump’s vacuum fitting. 6. Reconnect the vacuum pump to the mains power. PyroMark Q96 ID User Manual 01/2016 7-11...
  • Page 78 Maintenance This page intentionally left blank 7-12 PyroMark Q96 ID User Manual 01/2016...
  • Page 79 Glossary Troubleshooting If you need to contact QIAGEN Technical Services about an error, note down the steps leading to the error and any information given in any dialog boxes. This will help the QIAGEN Instrument Service Specialist in solving the problem.
  • Page 80 Check the sample and sample preparation. Contaminated samples will lead to an unusually high consumption of substrate at substrate addition, giving a high presequencing signal and no light produced at nucleotide dispensation. PyroMark Q96 ID User Manual 01/2016...
  • Page 81 Section 5.4. Reagents incorrectly stored or dissolved. Too much or too little template. Too much template DNA. Check amount of DNA template used. Reaction volume in wells was too high. Use lower volume. PyroMark Q96 ID User Manual 01/2016...
  • Page 82 Follow sample preparation instructions carefully, see Section 5.4. Dirty lens array. Clean the lens array, see Section 7.3. d) High baseline noise Contaminated nucleotides or background of contaminating sequence from PCR. Contact QIAGEN Technical Services if this persists. PyroMark Q96 ID User Manual 01/2016...
  • Page 83 Contact your system administrator. b) All lights on the Instrument software problem. Contact QIAGEN PyroMark Q96 ID Technical Services. remain on 5 minutes after it has been switched on PyroMark Q96 ID User Manual 01/2016...
  • Page 84 White remains If more than 1 min has elapsed since the plate (or (Sepharose beads) strips) was agitated, agitate again for 1 min in the before capturing the beads. immobilization plate PyroMark Q96 ID User Manual 01/2016...

  • Page 85: Glossary

    Instrument A method that describes physical settings for the parameters instrument, such as mixer frequency, block temperature, and pulse time settings. PyroMark Q96 ID User Manual 01/2016...
  • Page 86 Too high a substrate peak indicates that high levels of a contaminant might be present in, for example, buffers. Pyrogram The graph resulting from a sequencing reaction performed using Pyrosequencing technology. Each incorporated nucleotide is shown as a peak in Pyrogram. PyroMark Q96 ID User Manual 01/2016...
  • Page 87 SNPs and point mutations are structurally identical, polymorphism differing only in their frequency. Variations that occur in (SNP) 1% or less of a population are considered point mutations, while those occurring in more than 1% are SNPs. PyroMark Q96 ID User Manual 01/2016...
  • Page 88 Adenosine 5' phosphosulfate (APS) and Luciferin in the sequencing reaction. Theoretical The possible genotypes for a specific polymorphic position outcome are calculated on the basis of the sequence to analyze and the dispensation order. PyroMark Q96 ID User Manual 01/2016...

  • Page 89: Technical Data

    Appendix A Appendix A Technical data QIAGEN reserves the right to change specifications at any time. Environmental conditions Operating conditions Power 100–120 V, 220–240 V; 50/60 Hz Power consumption: 280 VA max. Overvoltage category Air temperature 18–28ºC (64–82ºF) Relative humidity 30–90% relative humidity (noncondensing) Altitude Up to 2000 m (6500 ft.)
  • Page 90 Appendix A Storage conditions Air temperature 10ºC to 40ºC (50ºF to 104ºF) Relative humidity Max. 75% (noncondensing) Mechanical data and hardware features: PyroMark Q96 ID Dimensions Width: 520 mm (20.47 in.) (closed) Height: 490 mm (19.29 in.) Depth: 620 mm (24.41 in.) Clearance Width: 540 mm (21.26 in.)
  • Page 91 200 MB Graphics card Supporting the resolution of the monitor Monitor 1280 x 800 pixels, True Color (32 bit) or higher Pointer device Mouse or similar Interfaces Ethernet ports (or external USB Ethernet adaptor). PyroMark Q96 ID User Manual 01/2016...

  • Page 92: Waste Electrical And Electronic Equipment (Weee

    Recycling can be provided by QIAGEN upon request at additional cost. In the European Union in accordance with the specific WEEE recycling requirements and, where a...

  • Page 93: Fcc Declaration

    Increase the separation between the equipment and  receiver. Connect the equipment into an outlet on a circuit  different from that to which the receiver is connected. Consult the dealer or an experienced radio/ TV  technician for help. PyroMark Q96 ID User Manual 01/2016...
  • Page 94 QIAGEN’s FCC Compliance Tests were conducted using QIAGEN-supported peripheral devices and shielded cables, such as those you receive with your system. Changes or modifications not expressly approved by QIAGEN could void the user’s authority to operate the equipment. PyroMark Q96 ID User Manual 01/2016...
  • Page 95 Appendix A Declaration of Conformity Name and address of the legal manufacturer QIAGEN GmbH QIAGEN Strasse 1 40724 Hilden Germany An up-to-date Declaration of Conformity can be requested from QIAGEN Technical Services. PyroMark Q96 ID User Manual 01/2016...
  • Page 96 Appendix A This page intentionally left blank PyroMark Q96 ID User Manual 01/2016...

  • Page 97: Assay Design And Validation

    The biotinylated primer should be purified by HPLC or an equivalent procedure since free biotin will compete with the biotinylated PCR product for binding sites on the streptavidin- coated Sepharose beads. PyroMark Q96 ID User Manual 01/2016...
  • Page 98 EDTA, and other chemical solvents than single-step enzyme-catalyzed processes. QIAGEN offers a complete range of nucleic acid preparation systems, ensuring the highest-quality templates for PCR. PyroMark Q96 ID User Manual 01/2016...
  • Page 99 PCR optimization The PyroMark PCR Kit will produce satisfactory results in most cases. However, if a higher Mg concentration is required, we recommend using the 25 mM MgCl provided in the kit. PyroMark Q96 ID User Manual 01/2016...
  • Page 100 (Cat. no. 59695), which provide bisulfite-treated completely methylated and unmethylated DNA in ready-to-use solutions. Regression analysis of the frequency of one allele measured in the PyroMark Q96 ID as a function of the input (expected) allele, should give an R value greater than 0.9.
  • Page 101 Never exclude the blank dispensations generated by the software as they function as excellent indicators of unspecific nucleotide incorporation. When manually generating a dispensation order, include an appropriate number of blank dispensations. If possible, start the dispensation order with PyroMark Q96 ID User Manual 01/2016...
  • Page 102 Ensure that the initial DNA sample is pure or that the assay is capable of specifically amplifying and/or sequencing only one target sequence in the sample. The assay may otherwise generate a mixed sequence that cannot be analyzed. PyroMark Q96 ID User Manual 01/2016...
  • Page 103 PCR product. This will show if the sequencing primer and the biotinylated primer can form duplexes and give background signal in Pyrosequencing reactions. Pyrograms from these controls should not show a peak after any nucleotide addition. PyroMark Q96 ID User Manual 01/2016...
  • Page 104 The analysis results for SQA assays are based on the appearance of the peaks in Pyrogram, related to peak height levels estimated by the software. The inclusion of known bases in SQA assays can improve the estimation of peak height level. PyroMark Q96 ID User Manual 01/2016...
  • Page 105 Appendix C Appendix C Liability clause QIAGEN shall be released from all obligations under its warranty in the event repairs or modifications are made by persons other than its own personnel, except in cases where the Company has given its written consent to perform such repairs or modifications.
  • Page 106 Appendix C This page intentionally left blank PyroMark Q96 ID User Manual 01/2016...
  • Page 107 Appendix D Appendix D Informations de sécurité Avant d’utiliser le PyroMark Q96 ID, il est impératif de lire attentivement ce manuel et de porter une attention particulière aux informations de sécurité. Afin de garantir un fonctionnement du système en toute sécurité et de maintenir le système en bon état de marche, il est impératif de suivre...
  • Page 108 AVERTISSEMENT/ Risque d’accident corporel et de détérioration ATTENTION du matériel L’appareil PyroMark Q96 ID est trop lourd pour être soulevé par une seule personne. Afin d’éviter tout accident corporel et toute détérioration du matériel, ne pas soulever l’appareil seul. PyroMark Q96 ID User Manual 01/2016...
  • Page 109 être sous tension et l’ouverture de capots ou le retrait de pièces risque d’exposer des éléments sous tension. Afin que le PyroMark Q96 ID fonctionne de manière satisfaisante et en toute sécurité, suivre les conseils suivants : Les cordons d’alimentation de l’appareil doivent être ...
  • Page 110 † ACGIH : American Conference of Government Industrial Hygienists (États-Unis d’Amérique) (Conférence américaine des hygiénistes industriels gouvernementaux). ‡ COSHH : Control of Substances Hazardous to Health (Royaume-Uni) (Contrôle des substances dangereuses pour la santé). PyroMark Q96 ID User Manual 01/2016...
  • Page 111 AVERTISSEMENT Pièces mobiles Afin d’éviter le contact avec les pièces en mouvement lorsque l’appareil PyroMark Q96 ID est en marche, son couvercle doit rester fermé. Ne pas démonter les panneaux du capot. Ils ne renferment aucune pièce réparable par l’utilisateur.
  • Page 112 Afin de garantir une bonne ventilation, laisser un dégagement d’au moins 10 cm sur les côtés et à l’arrière de l’appareil PyroMark Q96 ID. Les fentes et les ouvertures qui assurent la ventilation de l’appareil ne doivent pas être obstruées.
  • Page 113 à Chine (restriction de l’arrière de l’utilisation de certaines l’appareil substances dangereuses dans l’équipement électrique et électronique) Plaque Conforme aux normes signalétique à de sécurité des États l’arrière de Unis et du Canada l’appareil PyroMark Q96 ID User Manual 01/2016...
  • Page 114 Appendix D This page intentionally left blank PyroMark Q96 ID User Manual 01/2016...
  • Page 115 Sicherheitshinweise. Die Anweisungen und Sicherheitsinformationen in diesem Handbuch müssen vom Anwender befolgt werden, um einen sicheren Betrieb des PyroMark Q96 ID zu gewährleisten und das Gerät in einem sicheren Zustand zu erhalten. In diesem Handbuch werden die folgenden beiden...
  • Page 116 Wartung zurückzuführen sind. WARNUNG/ Verletzungsgefahr und Beschädigung des Geräts ACHTUNG Der PyroMark Q96 ID ist sehr schwer und sollte nicht von einer Person angehoben werden. Heben Sie das Gerät nicht allein an, um eine Verletzung und/oder Beschädi- gung des Geräts zu vermeiden.
  • Page 117 Öffnen der Abdeckungen oder das Entfernen von Gehäuseteilen können spannungsführende Komponenten freigelegt werden. Um einen zufriedenstellenden und sicheren Betrieb des PyroMark Q96 ID zu gewährleisten, befolgen Sie bitte die nachstehenden Hinweise: Die Geräte-Netzkabel müssen an Wechselstrom-  Steckdosen mit Schutzleiter (Erdungs-/Masseleiter) angeschlossen werden.
  • Page 118 * OSHA: Occupational Safety and Health Administration (Vereinigte Staaten von Amerika) † ACGIH: American Conference of Government Industrial Hygienists (Vereinigte Staaten von Amerika) ‡ COSHH: Control of Substances Hazardous to Health (Vereinigtes Königreich) PyroMark Q96 ID User Manual 01/2016...
  • Page 119 Appendix E Gefahren durch mechanische Teile Der Deckel des PyroMark Q96 ID muss während des Betriebs geschlossen sein. WARNUNG Sich bewegende Geräteteile Um einen Kontakt mit sich bewegenden Teilen beim Betrieb des PyroMark Q96 ID zu vermeiden, darf das Gerät nur mit geschlossenem Deckel betrieben werden.
  • Page 120 ID und der Wand eingehalten wird, damit eine ausreichende Belüftung des Geräts gewährleistet ist. Die Lüftungsschlitze und Öffnungen, die für die Be- und Entlüftung des PyroMark Q96 ID bestimmt sind, dürfen nicht verdeckt werden. Symbole auf dem PyroMark Q96 ID...
  • Page 121 Typenschild auf RCM-Mark-Zeichen für Australien Geräterückseite (Herstellerkennung: N17965) Typenschild auf Markierung gemäß RoHS-Richtlinie für Geräterückseite China (Beschränkung der Verwendung bestimmter gefährlicher Stoffe in Elektro- und Elektronikgeräten) Typenschild auf Entspricht US- amerikanischen und Geräterückseite kanadischen Sicherheitsstandards PyroMark Q96 ID User Manual 01/2016...
  • Page 122 Appendix E This page intentionally left blank PyroMark Q96 ID User Manual 01/2016...
  • Page 123 4-3 power requirements, 4-2 site requirements, 4-1 Safety Information, 1-1 Intended use, 2-3 chemical hazards, 1-3 electrical safety, 1-2 heat hazards, 1-4 mechanical hazards, 1-3 Liability clause, 1 proper use, 1-1 Sample preparation, 5-6 PyroMark Q96 ID User Manual 01/2016...
  • Page 124 Troubleshooting, 8-1 Sicherheitshinweise, 1 Tubing Software, 6-1 replacement, 7-10 data backup, 6-5 installation, 6-1 installation testing, 6-5 system requirements, 6-1 Ventilation, 1-4, 4-2 upgrading software, 6-2 Starting template, 2 Strand separation, 5-9 Waste disposal, 4 PyroMark Q96 ID User Manual 01/2016...
  • Page 126 Sample & Assay Technologies 1100446 01/2016...

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