Sample & Assay Technologies QIAsymphony® Virus/Bacteria
Contents
1.2. Pure viral bacterial nucleic acids 6 QlAsymphony Virus Bacteria Handbook 03 2011 Materials Provided Kit contents QlAsymphony Virus Bacteria Kit Mini 192 Midi 96 Catalog no 931036 931055 Number of preps 192 96 Reagent Cartridge 2 2 Enzyme Rack 2 2 Piercing Lid 2 2 Buffer AVE 20 ml 2 2 Buffer AVE 2 ml 2 2 Carrier RNA 2x1350 ug 2x1350 ug Reuse Seal Set 2 2 Handbook 1 Number of preps depends on the protocol used t Contains guanidine salts Not compatible with disinfectants containing bleach See page 9 for safety information Contains sodium azide as a preservative 8 A Reuse Seal Set contains 8 Reuse Seal Strips QlAsymphony Virus Bacteria Handbook 03 2011 7 Materials Required but Not Provided When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information please consult the appropriate material safety data sheets MSDSs available from the product supplier BM Sample Prep Cartridges 8 well cat no 997002 M 8 Rod Covers cat no 997004 BM Filter Tips 200 ul and 1500 ul cat nos 990332 and 997024 E Sample tubes see the relevant protocol sheet available at www qiagen com products qiasymphonvyvirus bacteriakits BM Vortexer For complex protocols M Buffer ATL cat no 939011 For using internal controls EM Sample tubes 14 ml 17 x 100 mm polystyrene round bottom tubes from Becton Dickins
3. drawer may lead to evaporation of eluates we strongly recommend using the cooling position Inventory scan Before starting a run the instrument checks that sufficient consumables for the queued batch es have been loaded into the corresponding drawers 14 QlAsymphony Virus Bacteria Handbook 03 2011 Preparation of sample material QlAsymphony Virus Bacteria Kits are suitable for use with a wide range of sample types including plasma serum CSF and respiratory and urogenital samples Prevent formation of foam in or on the samples Depending on the starting material sample pretreatment may be required Samples should be equilibrated to room temperature 15 25 C before starting the run For more information about the automated procedure including information about sample tubes that can be used with specific protocols and specific sample pretreatments see the relevant protocol sheet available at www qiagen com products qiasymphonyvirus bacteriakits Preparing carrier RNA Buffer AVE mixtures Note We strongly recommend use of carrier RNA If carrier RNA is not added recovery of nucleic acids may be significantly reduced To prepare a carrier RNA stock solution add 1350 pl Buffer AVE provided in 2 ml vials to the tube containing 1350 ug lyophilized carrier RNA to obtain a solution of 1 Ug ul Dissolve the carrier RNA thoroughly divide it into conveniently sized aliquots and store at 2 8 C for up to 2 weeks For vol
4. Technical 80 885942 Finland Orders 0800 914416 Fax 0800 914415 Technical 0800 914413 France Orders 01 60 920 926 Fax 01 60 920 925 Technical 01 60 920 930 Offers 01 60 920 928 Germany Orders 02103 29 12000 Fax 02103 29 22000 Technical 02103 29 12400 Hong Kong Orders 800 933 965 Fax 800 930 439 Technical 800 930 425 Ireland Orders 1800 555 049 Fax 1800 555 048 Technical 1800 555 061 Italy Orders 800 789 544 Fax 02 334304 826 Technical 800 787980 Japan Telephone 03 6890 7300 Fax 03 5547 0818 Technical 03 6890 7300 Korea South Orders 080 000 7146 Fax 02 2626 5703 Technical 080 000 7145 Luxembourg Orders 8002 2076 Fax 8002 2073 Technical 8002 2067 Mexico Orders 01 800 7742 639 Fax 01 800 1122 330 Technical 01 800 7742 436 The Netherlands Orders 0800 0229592 Fax 0800 0229593 Technical 0800 0229602 Norway Orders 800 18859 Fax 800 18817 Technical 800 18712 Singapore Orders 1800 742 4362 Fax 65 6854 8184 Technical 1800 742 4368 Spain Orders 91 630 7050 Fax 91 630 5145 Technical 91 630 7050 Sweden Orders 020 790282 Fax 020 790582 Technical 020 798328 Switzerland Orders 055 254 22 11 Fax 055 254 22 13 Technical 055 254 22 12 UK Orders 01293 422 911 Fax 01293 422 922 Technical 01293 422 999 USA Orders 800 426 8157 Fax 800 718 2056 Technical 800 DNA PREP 800 362 7737 QIAGEN A Sample amp Assay Technologies
5. R11 20 21 22 32 36 67 13 26 36 37 39 46 R11 Highly flammable R20 21 22 Harmful by inhalation in contact with skin and if swallowed R22 Harmful if swallowed R36 38 Irritating to eyes and skin R36 37 38 Irritating to eyes respiratory system and skin R42 43 May cause sensitization by inhalation and skin contact S7 Keep container tightly closed 13 Keep away from food drink and animal feedingstuffs S16 Keep away from sources of ignition No smoking 23 Do not breathe vapor 24 Avoid contact with the skin S26 In case of contact with eyes rinse immediately with plenty of water and seek medical advice 36 37 Irritating to eyes and respiratory system S36 37 39 Wear suitable protective clothing gloves and eye face protection 46 If swallowed seek medical advice immediately and show container or label QlAsymphony Virus Bacteria Handbook 03 2011 9 QSWT WX Contains guanidine hydrochloride and ethanol highly flammable harmful irritant Risk and safety phrases R11 22 36 38 S13 26 36 37 39 46 QSW5 Wil Contains guanidine hydrochloride and ethanol highly flammable harmful irritant Risk and safety phrases R1 1 22 36 38 13 26 36 37 39 46 QSW2 ew Contains ethanol highly flammable Risk and safety phrases R11 S7 16 Proteinase K X Contains proteinase K sensitizer irritant Risk and safety phrases R36 37 38 42 43 S23 24 26 36 37 24 hour emergency information Eme
6. March 2011 QlAsymphony Virus Bacteria Handbook QlAsymphony Virus Bacteria Mini Kit QlAsymphony Virus Bacteria Midi Kit For simultaneous purification of viral DNA and RNA from serum plasma or cerebrospinal fluid CSF and viral DNA viral RNA and bacterial DNA from respiratory and urogenital samples using the QlAsymphony SP QIAGEN Sample amp Assay Technologies QIAGEN Sample and Assay Technologies QIAGEN is the leading provider of innovative sample and assay technologies enabling the isolation and detection of contents of any biological sample Our advanced high quality products and services ensure success from sample to result QIAGEN sets standards in Purification of DNA RNA and proteins Nucleic acid and protein assays microRNA research and RNAi Automation of sample and assay technologies Our mission is to enable you to achieve outstanding success and breakthroughs For more information visit www giagen com Contents Intended Use 4 Summary and Explanation 4 Principles of the Procedure 4 Materials Provided 7 Kit contents 7 Materials Required but Not Provided 8 Warnings and Precautions 9 Reagent Storage and Handling 11 Specimen Handling and Storage 11 Procedure 12 Automated purification on the QlAsymphony SP 12 Protocol E General Purification Protocol 18 Troubleshooting Guide 22 Quality Control 25 Limitations 25 References 25 Contact Information 26 Ordering Information 27 QlAsymp
7. ation for internal controls and titer The protocol sheet also provides information for calculating the volume of internal control mixture according to the type of tube used We recommend preparing fresh mixtures for each run just before use Assay Control Sets Assay Control Sets are used for each protocol even when no internal controls are used A default Assay Control Set is preinstalled for each protocol Creation of additional Assay Control Sets is described in the QlAsymphony Management Console User Manual Note When using the default Assay Control Sets designed for working without internal control the use of carrier RNA Buffer AVE mixture is still required 16 QlAsymphony Virus Bacteria Handbook 03 2011 Handling RNA Ribonucleases RNases are very stable and active enzymes that generally do not require cofactors to function Since RNases are difficult to inactivate and only minute amounts are sufficient to destroy RNA do not use any plasticware or glassware without first eliminating possible RNase contamination Great care should be taken to avoid inadvertently introducing RNases into the RNA sample during or after the purification procedure Yields of nucleic acids Eluates prepared with carrier RNA may contain much more carrier RNA than target nucleic acids We recommend using quantitative amplification methods to determine yields Storing nucleic acids For short term storage of up to 24 hours we recommend storing purifi
8. bles drawer 8 Perform an inventory scan of the Reagents and Consumables drawer 9 Place the samples into the appropriate sample carrier and load them into the Sample drawer 10 Place the tube s containing the carrier RNA Buffer AVE mixture including optional internal control into the tube carrier and load into slot A of the Sample drawer For more information about preparing the mixture see the corresponding protocol sheet available at www qiagen com products qiasymphonyvirus bacteriakits as well as Preparing carrier RNA Buffer AVE mixtures page 15 and Using an internal control page 16 11 Using the touchscreen enter the required information for each batch of samples to be processed Enter the following information m Sample information depending on sample racks used m Protocol to be run Assay Control Set m Elution volume and output position m Tubes containing the carrier RNA Buffer AVE mixture including optional internal control 20 QlAsymphony Virus Bacteria Handbook 03 2011 After information about the batch has been entered the status changes from LOADED to QUEUED As soon as one batch is queued the Run button appears 12 Press the Run button to start the purification procedure All processing steps are fully automated At the end of the protocol run the status of the batch changes from RUNNING to COMPLETED 13 Retriev
9. d 8 Rod Covers used during a run are re racked in empty unit boxes in the Waste drawer Make sure that the Waste drawer contains sufficient empty unit boxes for plastic waste generated during the protocol run Note Ensure that the covers of the unit boxes are removed before loading the unit boxes into the Waste drawer If you are using 8 Rod Cover boxes for collecting used sample prep cartridges and 8 Rod Covers ensure that the box spacer has been removed A bag for used filter tips must be attached to the front side of the Waste drawer Note The presence of a tip disposal bag is not checked by the system Make sure that the tip disposal bag is properly attached before starting a protocol run For more information see the user manuals provided with your instrument Empty the tip bag after a maximum of 96 samples have been processed to avoid a tip jam A waste container collects liquid waste generated during the purification procedure The Waste drawer can only be closed if the waste container is in place Dispose of the liquid waste according to your local safety and environment regulations Do not autoclave the filled waste bottle Empty the waste bottle after a maximum of 96 samples have been processed Loading the Eluate drawer Load the required elution rack into the Eluate drawer Use Elution slot 1 with the corresponding cooling adapter As long term storage of eluates in the Eluate
10. e the elution rack containing the purified nucleic acids from the Eluate drawer For short term storage of up to 24 hours we recommend storing purified nucleic acids at 2 8 C For long term storage of over 24 hours we recommend storage purified nucleic acids at 20 C We recommend removing the eluate plate from the Eluate drawer immediately after the run has finished Depending on temperature and humidity elution plates left in the QlAsymphony SP after the run is completed may experience condensation or evaporation Result files are generated for each elution plate Note Ensure that the correct eluate volume is used for the downstream applications 14 If a reagent cartridge is only partially used seal it with the provided Reuse Seal Strips and close tubes containing proteinase K with screw caps immediately after the end of the protocol run to avoid evaporation If Buffer ATL was used close the bottle and store at 15 25 C Note For more information about storage of partially used reagent cartridges see Storage page 11 15 Discard used sample tubes plates and waste according to your local safety regulations See page 9 for safety information 16 Clean the QlAsymphony SP Follow the maintenance instructions in the user manuals supplied with your instrument Make sure to clean the tip guards regularly to minimize the risk of cross contamination 17 Close the instrument drawers and switch off the QlAs
11. ed nucleic acids at 2 8 C For long term storage of over 24 hours we recommend storage at 20 C QlAsymphony Virus Bacteria Handbook 03 2011 17 Protocol General Purification Protocol The following is a general protocol for using QlAsymphony Virus Bacteria Kits Detailed information for each protocol including volumes and tubes is provided in protocol sheets that can be downloaded at www qiagen com products qiasymphonyvirus bacteriakits Important points before starting M Ensure that you are familiar with operating the QlAsymphony SP Refer to the user manuals supplied with your instrument for operating instructions E Optional maintenance is not mandatory for instrument function but is highly recommended to reduce risk of contamination M Ensure you are familiar with the protocol sheet corresponding to the procedure you want to use available at www qiagen com products qiasymphonyvirus bacteriakits In particular note the initial elution volumes required for accurate calculation of internal controls and titer as well as instructions for calculating the volume of internal control mixture according to the type of tube used Also check if the protocol requires Buffer ATL M Before using a reagent cartridge for the first time check that Buffers QSL2 and QSB1 do not contain a precipitate If necessary remove the troughs containing Buffers QSL2 and QSBT from the reagent cartridge and incubate for 30 minutes at 37 C with occas
12. er s responsibility to validate system performance for any procedures used in their laboratory that are not covered by the QIAGEN performance evaluation studies References QIAGEN maintains a large up to date online database of scientific publications utilizing QIAGEN products Comprehensive search options allow you to find the articles you need either by a simple keyword search or by specifying the application research area title etc For a complete list of references visit the QIAGEN Reference Database online at www qiagen com RefDB search asp or contact QIAGEN Technical Services or your local distributor QlAsymphony Virus Bacteria Handbook 03 2011 25 Contact Information At QIAGEN we pride ourselves on the quality and availability of our technical support Our Technical Service 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 QlAsymphony Virus Bacteria Mini Kit QlAsymphony Virus Bacteria Midi Kit or QIAGEN products in general please 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 perfo
13. h is located at the bottom left corner of the QlAsymphony SP Log in to the instrument gt 4 Ensure the Waste drawer is prepared properly and perform an inventory scan of the Waste drawer including the tip chute and liquid waste Replace the tip disposal bag if necessary 5 Load the required elution rack into the Eluate drawer QlAsymphony Virus Bacteria Handbook 03 2011 19 Use only Elution slot 1 with the corresponding cooling adapter When using a 96 well plate make sure that the plate is in the correct orientation as incorrect placement may cause sample mixup in downstream analysis 6 Load the required reagent cartridge s and consumables into the Reagents and Consumables drawer 7 If using a protocol that does not require Buffer ATL continue with step 8 If using a protocol that requires Buffer ATL press the R C button in the touchscreen to open the screen that shows the consumables status Consumables 8 Rod Covers Tubes Filter Tips Reagent Cartridges Press the Scan Bottle button to scan the bar code of the bottle of Buffer ATL with the handheld bar code scanner Press OK Ensure that the bottle of Buffer ATL is scanned opened and placed into the position specified in the touchscreen before starting the inventory scan Otherwise the inventory scan must be repeated after scanning opening and placing the bottle of Buffer ATL into the Reagents and Consuma
14. he Reuse Seal Strips have been removed M Before starting the procedure ensure that the magnetic particles are fully resuspended Vortex the trough containing the magnetic particles vigorously for at least 3 minutes before first use M Before loading the reagent cartridge remove the cover from the trough containing the magnetic particles and open the enzyme tubes Make sure that the enzyme has been equilibrated to room temperature 15 25 C Make sure that the piercing lid is placed on the reagent cartridge or if using a partially used reagent cartridge make sure the Reuse Seal Strips have been removed M If samples are bar coded orient samples in the tube carrier so that the bar codes face the bar code reader at the left side of the QlAsymphony SP BM For information about sample tubes compatible with a certain protocol see the corresponding protocol sheet available at www qiagen com products qiasymphonvyvirus bacteriakits BM For information about minimum sample volumes for samples in primary and secondary tubes for a certain protocol see the corresponding protocol sheet available at www qiagen com products qiasymphonvyvirus bacteriakits This information also indicates which tubes can be used for the different protocols Procedure 1 Close all drawers and the hood 2 Switch on the QlAsymphony SP and wait until the Sample Preparation screen appears and the initialization procedure has finished The power switc
15. he troughs are reclosed with Reuse Seal Strips and incubate the complete reagent cartridge for 30 min at 37 C with occasional shaking in a water bath Ensure that instruments have been checked maintained and calibrated regularly according to the manufacturer s instructions DEREK WWO _ _ _ _ _ _ _ _ _ __ QlAsymphony Virus Bacteria Handbook 03 2011 23 Comments and suggestions f Clogging of pipet tip Insoluble material was not removed from the due to insoluble sample prior to starting the QlAsymphony material purification procedure To remove insoluble material for viral applications centrifuge the sample at 3000 x g for 1 min and transfer the supernatant to a new sample tube If required use pretreatment procedures as described in the corresponding protocol sheets for example for viscous sample materials Protocol sheets are available at www qiagen com products qiasymphonvyvirus bacteriakits weie a ae Z O Z ER 24 QlAsymphony Virus Bacteria Handbook 03 2011 Quality Control In accordance with QIAGEN s ISO certified Quality Management System each lot of QlAsymphony Virus Bacteria Kit is tested against predetermined specifications to ensure consistent product quality Limitations QlAsymphony Virus Bacteria Kits are intended for molecular biology applications These products are not intended for the diagnosis prevention or treatment of a disease It is the us
16. hen stored properly the kit is stable until the expiration date on the kit box Partially used reagent cartridges can be stored for a maximum of 2 weeks enabling cost efficient reuse of reagents and more flexible sample processing If a reagent cartridge is partially used replace the cover of the trough containing the magnetic particles and seal the reagent cartridge with the provided Reuse Seal Strips immediately after the end of the protocol run to avoid evaporation Running batches with low sample numbers lt 24 will potentially reduce the total number of sample preparations possible per cartridge To avoid reagent evaporation the reagent cartridge should be open for a maximum of 15 hours including run times at a maximum environmental temperature of 30 C Avoid exposure of the reagent cartridges to UV light e g used for decontamination as exposure may cause accelerated aging of the reagent cartridges RC and buffers Note The label on the QlAsymphony Virus Bacteria Kit box displays the expiration date of the kit The result file documents the expiration dates for only the reagent cartridge RC and Buffer ATL if required Specimen Handling and Storage Prevent formation of foam in or on the samples Depending on the starting material sample pretreatment may be required Samples should be equilibrated to room temperature 15 25 C before starting the run For more information about the automated procedure including
17. hony Virus Bacteria Handbook 03 2011 3 Intended Use QlAsymphony Virus Bacteria Kits are intended to be used only in combination with the QlAsymphony SP Summary and Explanation QlAsymphony Virus Bacteria Kits provide reagents for fully automated and simultaneous purification of viral nucleic acids from serum plasma or CSF or viral nucleic acids and bacterial DNA from various materials including respiratory samples such as swabs aspirates sputum bronchoalveolar lavage BAL as well as urine and urogenital swabs cervical and urethral The kits can be used to purify nucleic acids from a broad range of DNA and RNA viruses as well as bacterial DNA from Gram negative and Gram positive bacteria Proven performance leading magnetic particle technology enables purification of high quality nucleic acids that are free of proteins nucleases and other impurities The purified nucleic acids are ready for direct use in downstream applications such as amplification or other enzymatic reactions The QlAsymphony SP performs all steps of the purification procedure Up to 96 samples in batches of up to 24 are processed in a single run Principles of the Procedure QlAsymphony technology combines the speed and efficiency of silica based nucleic acid purification with the convenient handling of magnetic particles Figure 1 The purification procedure is designed to ensure safe and reproducible handling of potentially infectious samples and compr
18. iagen com Other than expressly stated licenses QIAGEN makes no warranty that these Kits and or their use s do not infringe the rights of third parties These Kits and their components are licensed for one time use and may not be reused refurbished or resold QIAGEN specifically disclaims any other licenses expressed or implied other than those expressly stated OW RY ZO R The purchaser and user of the Kits agree not to take or permit anyone else to take any steps that could lead to or facilitate any acts prohibited above QIAGEN may enforce the prohibitions of this Limited License Agreement in any Court and shall recover all its investigative and Court costs including attorney fees in any action to enforce this Limited License Agreement or any of its intellectual property rights relating to the Kits and or their components For updated license terms see www qiagen com 2008 11 QIAGEN all rights reserved www qiagen com Australia Orders 1 800 243 800 Fax 03 9840 9888 Technical 1 800 243 066 Austria Orders 0800 28 10 10 Fax 0800 28 10 19 Technical 0800 28 10 11 Belgium Orders 0800 79612 Fax 0800 79611 Technical 0800 79556 Brazil Orders 0800 557779 Fax 55 11 5079 4001 Technical 0800 557779 Canada Orders 800 572 9613 Fax 800 713 5951 Technical 800 DNA PREP 800 362 7737 China Orders 86 21 3865 3865 Fax 86 21 3865 3965 Technical 800 988 0325 Denmark Orders 80 885945 Fax 80 885944
19. idges Nonsterile polypropylene tubes 0 85 ml maximum capacity less than 0 7 ml storage capacity 0 4 ml elution capacity 2304 in racks of 96 includes cap strips QlAsymphony sample prep module year warranty on parts and labor Cat no 9018087 9018088 9241032 9241033 9241034 9018577 990332 997024 997006 19588 9001297 For up to date licensing information and product specific disclaimers see the respective QIAGEN kit handbook or user manual QIAGEN kit handbooks and user manuals are available at www qiagen com or can be requested from QIAGEN Technical Services or your local distributor 28 QlAsymphony Virus Bacteria Handbook 03 2011 Notes QlAsymphony Virus Bacteria Handbook 03 2011 29 Notes 30 QlAsymphony Virus Bacteria Handbook 03 2011 Trademarks QIAGEN QlAsymphony QIAGEN Group Limited License Agreement Use of this product signifies the agreement of any purchaser or user of the QlAsymphony Virus Bacteria Kits to the following terms 1 The QlAsymphony Virus Bacteria Kits may be used solely in accordance with the QlAsymphony Virus Bacteria Handbook and for use with components contained in the Kits only QIAGEN grants no license under any of its intellectual property to use or incorporate the enclosed components of these Kits with any components not included within these Kits except as described in the QlAsymphony Virus Bacteria Handbook and additional protocols available at www q
20. information about sample tubes that can be used with specific protocols and specific sample pretreatments see the relevant protocol sheet available at www qiagen com products qiasymphonyvirus bacteriakits zn _ _ _ _ _ _ LL QlAsymphony Virus Bacteria Handbook 03 2011 11 Procedure Automated purification on the QlAsymphony SP The QlAsymphony SP makes automated sample preparation easy and convenient Samples reagents and consumables and eluates are separated in different drawers Simply load samples reagents provided in special cartridges and preracked consumables in the appropriate drawer before a run Start the protocol and remove purified DNA from the Eluate drawer after processing Refer to the user manuals supplied with your instrument for operating instructions Note Optional maintenance is not mandatory for instrument function but is highly recommended to reduce risk of contamination The range of protocols available is continually expanding and additional QIAGEN protocols can be downloaded free of charge at www qiagen com products giasymphonyvirus bacteriakits Loading reagent cartridges into the Reagents and Consumables drawer Reagents for purification of DNA are contained in an innovative reagent cartridge see Figure 2 Each trough of the reagent cartridge contains a particular reagent such as magne
21. ional shaking to dissolve precipitate Make sure to replace the troughs in the correct positions If the reagent cartridge is already pierced make sure that the troughs are sealed with Reuse Seal Strips and incubate the complete reagent cartridge for 30 minutes at 37 C with occasional shaking in a water bath BM Try to avoid vigorous shaking of the reagent cartridge otherwise foam may be generated which can lead to liquid level detection problems M Before starting a protocol that requires Buffer ATL check whether precipitate has formed in Buffer ATL If necessary dissolve by heating at 70 C with gentle agitation in a water bath Aspirate bubbles from the surface of Buffer ATL Things to do before starting M Prepare all required mixtures including mixtures containing carrier RNA and internal controls optional just before starting For more information see the corresponding protocol sheet available at Ensure that instruments have been checked maintained and calibrated regularly according to the manufacturer s instructions 18 QlAsymphony Virus Bacteria Handbook 03 2011 www qiagen com products qiasymphonyvirus bacteriakits as well as Preparing carrier RNA Buffer AVE mixtures page 15 and Using an internal control page 16 M Make sure that the piercing lid is placed on the reagent cartridge and the lid of the magnetic particle trough has been removed or if using a partially used reagent cartridge make sure t
22. ises 4 steps lyse bind wash and elute see flowchart page 6 The user can choose between different elution volumes Number of preps depends on the sample volume and protocol used 4 QlAsymphony Virus Bacteria Handbook 03 2011 Magnetic P rod Slow up and down movement to collect magnetic particles Rod cover Magnetic q rod Fast up and down movement of rod cover to release magnetic particles Reagent 2 and magnetic particles Reagent 1 and magnetic particles A Well 2 Figure 1 Schematic of the QlAsymphony SP principle The QlAsymphony SP processes a sample containing magnetic particles as follows A magnetic rod protected by a rod cover enters a well containing the sample and attracts the magnetic particles The magnetic rod cover is positioned above another well and the magnetic particles are released The QlAsymphony SP uses a magnetic head containing an array of 24 magnetic rods and can therefore process up to 24 samples simultaneously Steps 1 and 2 are repeated several times during sample processing SEE KW ROK QlAsymphony Virus Bacteria Handbook 03 2011 5 QlAsymphony Virus Bacteria Procedure Sample Lysis Lysate and magnetic particles transferred to sample prep cartridge Viral bacterial nucleic acids bind to magnetic particles Magnetic separation dS Auoyduiksyjp s uo uo D gt IJIINd poo d1a 9NU payoWoynD AJJN4 Magnetic separation Elute
23. on cat no 352051 www bd com BM Sample tubes 2 ml sample tubes with screw caps or without screw caps from Sarstedt e g cat nos 72 693 and 72 694 www sarstedt com 8 QlAsymphony Virus Bacteria Handbook 03 2011 Warnings and Precautions When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information please consult the appropriate material safety data sheets MSDSs These are available online in convenient and compact PDF format at www giagen com Support MSDS aspx where you can find view and print the MSDS for each QIAGEN kit and kit component CAUTION DO NOT add bleach or acidic solutions directly to the sample preparation waste Buffers in the reagent cartridge RC contain guanidine salts which can form highly reactive compounds when combined with bleach If liquid containing these buffers is spilt clean with suitable laboratory detergent and water If the spilt liquid contains potentially infectious agents clean the affected area first with laboratory detergent and water and then with 1 v v sodium hypochlorite The following risk and safety phrases apply to components of QlAsymphony Virus Bacteria Kits QSL2 X Contains guanidine thiocyanate harmful Risk and safety phrases R20 21 22 32 13 26 36 37 39 46 QSB1 WX Contains isopropanol and guanidine thiocyanate highly flammable harmful irritant Risk and safety phrases
24. oplates MTP For use in the QlAsymphony Eluate drawer Cooling adapter for EMT racks For use in the QlAsymphony Eluate drawer QlAsymphony Virus Bacteria Handbook 03 2011 Cat no 931036 931055 939011 19131 19133 997002 997004 997008 997012 9013395 9017660 9018085 9018086 27 Product Cooling Adapter PCR Qsym Cooling Adapter tubes 2 ml Qsym Tube Insert 2 ml sample carrier Qsym Tube Insert 11 mm sample carrier Qsym Tube Insert 13 mm sample carrier Qsym Adapter tubes 2 ml Qsym Filter Tips 200 ul 1024 Filter Tips 1500 ul 1024 Reuse Seal Set 20 Elution Microtubes CL 24 x 96 QlAsymphony SP Contents Cooling adapter for PCR plates For use in the QlAsymphony Eluate drawer Cooling adapter for 2 ml screw cap tubes For use in the QlAsymphony Eluate drawer Secondary tube adapter for 2 ml screw cap tubes for use with the QlAsymphony tube carrier Primary tube adapter 11 mm for use with the QlAsymphony tube carrier Primary tube adapter 13 mm for use with the QlAsymphony tube carrier Adapter for 2 ml screw cap tubes For use in the QlAsymphony Eluate drawer Disposable Filter Tips racked 8 x 128 For use with the QlAcube and the QlAsymphony SP Disposable Filter Tips racked 8 x 128 For use with the QlAsymphony SP Reuse seal sets for sealing partly used QlAsymphony reagent cartr
25. rgency medical information in English French and German can be obtained 24 hours a day from Poison Information Center Mainz Germany Tel 49 6131 19240 R11 Highly flammable R20 21 22 Harmful by inhalation in contact with skin and if swallowed R22 Harmful if swallowed R36 38 Irritating to eyes and skin R36 37 38 Irritating to eyes respiratory system and skin R42 43 May cause sensitization by inhalation and skin contact S7 Keep container tightly closed 13 Keep away from food drink and animal feedingstuffs S16 Keep away from sources of ignition No smoking 23 Do not breathe vapor 24 Avoid contact with the skin S26 In case of contact with eyes rinse immediately with plenty of water and seek medical advice 36 37 Irritating to eyes and respiratory system 36 37 39 Wear suitable protective clothing gloves and eye face protection 46 If swallowed seek medical advice immediately and show container or label 10 QlAsymphony Virus Bacteria Handbook 03 2011 Reagent Storage and Handling The QlAsymphony Virus Bacteria Kits should be stored at room temperature 15 25 C The magnetic particles in the reagent cartridges remain active when stored at this temperature Do not store reagent cartridges at temperatures below 15 C Store lyophilized carrier RNA and Buffer AVE at room temperature QlAsymphony Virus Bacteria Kits contain ready to use proteinase K solution that can be stored at room temperature W
26. rmance or new applications and techniques For technical assistance and more information please see our Technical Support Center at www qiagen com Support or call one of the QIAGEN Technical Service Departments or local distributors see back cover or visit www qiagen com p k 26 QlAsymphony Virus Bacteria Handbook 03 2011 Ordering Information Product QlAsymphony Virus Bacteria Mini Kit 192 QlAsymphony Virus Bacteria Midi Kit 96 Related products Buffer ATL 4 x 50 ml QIAGEN Proteinase K 2 ml QIAGEN Proteinase K 10 ml Sample Prep Cartridges 8 well 336 8 Rod Covers 144 Reagent Cartridge Holder 2 Accessory Trough 10 Tip Disposal Bags 15 Sample Carrier plate Qsym Cooling Adapter MTP RB Qsym Cooling Adapter EMT Qsym Contents Includes 2 reagent cartridges and enzyme racks and accessories Includes 2 reagent cartridges and enzyme racks and accessories Buffer 4 x 50 ml ATL for use with QlAsymphony complex protocols 2 ml gt 600 mAU ml solution 10 ml gt 600 mAU mil solution 8 well sample prep cartridges for use with the QlAsymphony SP 8 Rod Covers for use with the QlAsymphony SP Reagent cartridge holder for use with the QlAsymphony SP Accessory troughs for use with the QlAsymphony SP Tip disposal bags for use with the QlAsymphony SP Plate carrier for sample input for use with the QlAsymphony SP Cooling adapter for round bottom micr
27. ted slots see Figure 2 above the reagent cartridge is subsequently loaded into the Reagents and Consumables drawer Piercing lid Figure 3 Easy worktable setup with reagent cartridges Partially used reagent cartridges can be stored until needed again see Storage page 11 Loading plasticware into the Reagents and Consumables drawer Sample prep cartridges 8 Rod Covers both preracked in unit boxes and disposable filter tips 200 ul tips provided in blue racks 1500 ul tips provided in gray racks are loaded into the Reagents and Consumables drawer Note Ensure that the covers of the unit boxes are removed before loading the unit boxes into the Reagents and Consumables drawer Note Tips have filters to help prevent cross contamination QlAsymphony Virus Bacteria Handbook 03 2011 13 Tip rack slots on the QlAsymphony SP worktable can be filled with either type of tip rack The QlAsymphony SP will identify the type of tips loaded during the inventory scan Note Do not refill tip racks or unit boxes for sample prep cartridges or 8 Rod Covers before starting another protocol run The QlAsymphony SP can use partially used tip racks and unit boxes For the consumables required see the relevant protocol sheet available at www qiagen com products qiasymphonyvirus bacteriakits For plasticware ordering information see page 27 Loading the Waste drawer Sample prep cartridges an
28. tic particles lysis buffer wash buffer or elution buffer Partially used reagent cartridges can be reclosed with Reuse Seal Strips for later reuse which avoids generation of waste due to leftover reagents at the end of the purification procedure Reuse Seal Strip _AV 4 Magnetic particle moss PZ trough Frame with 1 S reagent troughs R Reagent cartridge holder Piercing lid Enzyme rack Slots for screw caps from enzyme tubes Figure 2 QlAsymphony reagent cartridge The reagent cartridge contains all reagents required for the protocol run 12 QlAsymphony Virus Bacteria Handbook 03 2011 Before starting the procedure ensure that the magnetic particles are fully resuspended Remove the magnetic particle trough from the reagent cartridge frame vortex it vigorously for at least 3 minutes and replace it in the reagent cartridge frame before the first use Place the reagent cartridge into the reagent cartridge holder Place the empty enzyme rack into the reagent cartridge holder Before using a reagent cartridge for the first time place the piercing lid PL on top of the reagent cartridge Figure 3 Note The piercing lid is sharp Take care when placing it onto the reagent cartridge Make sure to place the piercing lid onto the reagent cartridge in the correct orientation After the magnetic particle trough cover is removed and the enzyme rack tubes are opened screw caps can be stored in dedica
29. tor the efficiency of sample preparation and downstream assay Internal controls must be added with carrier RNA Buffer AVE mixture and the total volume of the internal control carrier RNA Buffer AVE mixture remains 120 ul The amount of internal control added depends on the assay system and the elution volume chosen within the QlAsymphony SP protocol Calculation and validation must be performed by the user Refer to the manufacturer s instructions for the downstream assay to determine the optimal concentration of internal control Using a concentration other than that recommended may lead to incorrect results especially if the internal control is used for calculation of titers A mixture of internal controls can be used to analyze different parameters from a single eluate Compatibility of different internal controls must be validated by the user When calculating the amount of internal control to use as well as the titer of the processed sample it is necessary to take into consideration the actual volume of elution solution that is used for each sample Since small amounts of liquid are lost during transfer and contact with the magnetic particles the initial volume of elution solution must be larger than the selected volume to ensure that the final eluate is of the correct volume The relevant protocol sheet available at www qiagen com products qiasymphonyvirus bacteriakits provides the initial elution volumes to allow accurate calcul
30. umes of carrier RNA required for specific protocols see the relevant protocol sheet available at www qiagen com products qiasymphonyvirus bacteriakits Calculating the volume of carrier RNA mixture per tube The minimum volume of carrier RNA Buffer AVE mixture must include sufficient additional volume to take into account liquid loss due to pipetting and evaporation Compatible tube formats including minimum volume of carrier RNA Buffer AVE mixtures are listed at www qiagen com products qiasymphonyvirus bacteriakits Tubes containing carrier RNA Buffer AVE mixtures are placed in a tube carrier The tube carrier containing the carrier RNA Buffer AVE mixture s must be placed in slot A of the sample drawer Up to 8 tubes of the mixture can be used per batch and up to 24 tubes can be used per run of 4 batches If less carrier RNA has been shown to be better for your amplification system adjust the volume of carrier RNA accordingly The use of a different concentration of carrier RNA must be validated for each particular sample type and downstream assay If no carrier RNA is used the tubes loaded into slot A must contain Buffer AVE only 120 ul Buffer AVE per sample QlAsymphony Virus Bacteria Handbook 03 2011 15 Using an internal control Using QlAsymphony Virus Bacteria Kits in combination with amplification systems that use an internal control may require introduction of these internal controls into the purification procedure to moni
31. y pierced make sure that the trough is reclosed with a Reuse Seal Strip and incubate the complete reagent cartridge in a water bath at 37 C for 30 min with occasional shaking Ensure that instruments have been checked maintained and calibrated regularly according to the manufacturer s instructions 22 QlAsymphony Virus Bacteria Handbook 03 2011 Comments and suggestions Low yield of nucleic acids a Magnetic particles were Before starting the procedure ensure that the not completely magnetic particles are fully resuspended Vortex resuspended for at least 3 min before use b Frozen samples were Thaw frozen samples with mild agitation to not mixed properly ensure thorough mixing after thawing c Carrier RNA not added Reconstitute carrier RNA in Buffer AVE and mix with appropriate volume of Buffer AVE as described in Preparing carrier RNA Buffer AVE mixtures starting on page 15 Repeat the purification procedure with new samples Degraded nucleic acids Samples were stored incorrectly or subjected to too many freeze thaw cycles Repeat the purification procedure with new samples e Incomplete sample lysis Before use check that Buffer QSL2 and QSB1 do not contain precipitates If necessary remove the troughs containing Buffers QSL2 and QSB1 from the reagent cartridge and incubate for 30 min at 37 C with occasional shaking to dissolve precipitate If the reagent cartridge is already pierced make sure that t
32. ymphony SP QlAsymphony Virus Bacteria Handbook 03 2011 21 Troubleshooting Guide This troubleshooting guide may be helpful in solving any problems that may arise For more information see also the Frequently Asked Questions page at our Technical Support Center www qiagen com FAQ FAQList aspx The scientists in QIAGEN Technical Services are always happy to answer any questions you may have about either the information and protocols in this handbook or sample and assay technologies for contact information see back cover or visit www qiagen com Comments and suggestions General handling Error message If an error message is displayed during a displayed in the protocol run refer to the user manuals supplied touchscreen with your instrument Precipitate in reagent trough of opened cartridge a Buffer evaporation Excessive evaporation may lead to increased salt concentration in buffers Discard reagent cartridge Make sure to seal buffer troughs of a partially used reagent cartridge with Reuse Seal Strips when not being used for purification b Storage of reagent Storage of reagent cartridge under 15 C may cartridge lead to formation of precipitates If necessary remove the troughs containing Buffer QSL2 and QSB1 from the reagent cartridge and incubate in a water bath at 37 C for 30 min with occasional shaking to dissolve precipitate Make sure to replace the trough in the correct position If the reagent cartridge is alread
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