QIAamp MinElute Virus Vacuum Handbook
Contents
1. reduced hands on time in purification procedures In combination with the QlAvac Connecting System optional the QlAvac 24 Plus can be used as a flow through system The sample flow through is collected in a separate waste bottle For maintenance of the QlAvac 24 Plus please refer to the handling guidelines in the QlAvac 24 Plus Handbook Processing GlAamp MinElute columns on the QlAvac 24 Plus GlAamp MinElute spin columns are processed on the QlAvac 24 Plus using disposable VacConnectors and reusable VacValves VacValves optional are inserted directly into the luer slots of the QlAvac 24 Plus manifold and ensure a steady flow rate facilitating parallel processing of samples of different natures e g serum and plasma volumes or viscosities They should be used if sample flow rates differ significantly in order to ensure consistent vacuum VacConnectors are disposable connectors that fit between GlAamp MinElute columns and VacValves or between the GlAamp MinElute columns and the luer slots of the QlAvac 24 Plus They prevent direct contact between the spin column and VacValve during purification thereby avoiding any cross contamination between samples VacConnectors are discarded after a single use 14 QlAamp MinElute Virus Vacuum Handbook 04 2010 Handling guidelines for the AlAvac 24 Plus Always place the QlAvac 24 Plus on a secure bench top or work area If dropped the QlAvac 24 Plus manifold may crack Always store the2. AW2 prepared with 70 ethanol i Protease Resuspension Buffer used with incompatible starting materials If tubes containing Buffer AVE are accessed repeatedly be careful to not introduce RNases In case of RNase contamination replace the open vial of Buffer AVE with a new vial Repeat the purification procedure with new samples Check that Buffers AW1 and AW2 concentrates were diluted with the correct volume of ethanol Repeat the purification procedure with new samples Check that Buffer AW1 and AW2 concentrates were diluted with 96 100 ethanol Do not use denatured alcohol which contains other substances such as methanol or methylethylketone Repeat the purification procedure with new samples Protease Resuspension Buffer is not compatible with samples or internal controls that contain phosphate e g viral transport medium cell culture supernatants or phosphate buffered saline If the sample or internal control contains phosphate it is highly recommended to resuspend QIAGEN Protease in Buffer AVE See Preparation of QIAGEN Protease page 20 RNA or DNA does not perform well in downstream enzymatic reactions a Little or no RNA in the eluate b Samples frozen and thawed more than once c Low concentration of virus in the samples d Insufficient sample lysis in Buffer AL See Little or no nucleic acid in the eluate for possible reasons Increase the amount of eluate added to the reaction if possibl
3. If carrier RNA is not added to Buffer AL this may lead to reduced viral RNA or DNA recovery The amount of lyophilized carrier RNA provided is sufficient for the volume of Buffer AL supplied with the kit The concentration of carrier RNA has been adjusted so that the QlAamp MinElute Virus Vacuum protocol can be used as a generic purification system compatible with many different amplification systems and is suitable for a wide range of RNA and DNA viruses Different amplification systems vary in efficiency depending on the total amount of nucleic acid present in the reaction Eluates from this kit contain both viral nucleic acids and carrier RNA and amounts of carrier RNA will greatly exceed amounts of viral nucleic acids Calculations of how much eluate to add to downstream amplifications should therefore be based on the amount of carrier RNA added To obtain the highest levels of sensitivity in amplification reactions it may be necessary to adjust the amount of carrier RNA added to Buffer AL 10 QlAamp MinElute Virus Vacuum Handbook 04 2010 The GlAamp MinElute Virus Vacuum Procedure Sample I Lyse Bind SED Vacuum M Wash Buffer AW1 Remove extender before vacuum is switched on KD Vacuum Wash Buffer AW2 SEED Vacuum Wash I ethanol Dry spin Elute Pure viral nucleic acids QlAamp MinElute Virus Vacuum Handbook 04 2010 11 Addition of internal controls Usin
4. 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 m Sample amp Assay Technologies
5. 00 ml of the solution to be treated and shake vigorously to bring the DEPC into solution or let the solution incubate for 12 hours at 37 C Autoclave for 15 minutes to remove any trace of DEPC It may be desirable to test water sources for the presence of contaminating RNases since many sources of distilled water are free of RNase activity Note GlAamp MinElute Virus Vacuum Kit buffers are not rendered RNase free by DEPC treatment and are therefore free of any DEPC contamination When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information consult the appropriate material safety data sheets MSDSs available from the product supplier Plastics used for some electrophoresis tanks are not resistant to ethanol Take proper care and check the supplier s instructions QlAamp MinElute Virus Vacuum Handbook 04 2010 31 Ordering Information Product Contents Cat no QlAamp MinElute Virus Vacuum For 50 minipreps 50 GlAamp 57714 Kit 50 MinElute Columns QIAGEN Protease Carrier RNA Buffers Extension Tubes 3 ml Collection Tubes 1 5 ml Accessories QlAvac 24 Plus Vacuum Manifold for processing 19413 1 24 spin columns includes QlAvac 24 Plus Vacuum Manifold Luer Plugs Quick Couplings VacValves 24 24 valves for use with the QlAvac 19408 24 Plus VacConnectors 500 500 disposable connectors for use 19407 with QlAamp spin columns on luer slots or VacValves QlA
6. 1 5 ml microcentrifuge tubes provided If the purified viral RNA and DNA is to be stored for up to 24 hours storage at 2 8 C is recommended For periods of storage longer than 24 hours storage at 20 C is recommended QlAamp MinElute Virus Vacuum Handbook 04 2010 9 Yield and size of viral nucleic acids Yields of viral nucleic acid isolated from biological samples are normally below 1 pg and are therefore difficult to determine with a spectrophotometer Quantitative amplification methods are recommended for determination of yields When quantifying nucleic acids isolated using the QlAamp MinElute Virus Vacuum protocol remember that there will be much more carrier RNA in the sample than viral RNA The size distribution of viral nucleic acid purified using this procedure can be checked by agarose gel electrophoresis and hybridization to a virus specific labeled probe followed by autoradiography Sambrook J and Russell D W 2001 Molecular Cloning A Laboratory Manual 3rd ed Cold Spring Harbor NY Cold Spring Harbor Laboratory Press Carrier RNA Carrier RNA serves two purposes Firstly it enhances binding of viral nucleic acids to the QlAamp MinElute membrane especially if there are very few target molecules in the sample Secondly the addition of large amounts of carrier RNA reduces the chance of viral RNA degradation in the rare event that RNase molecules escape denaturation by the chaotropic salts and detergent in Buffer AL
7. 19413 and a vacuum pump capable of producing a vacuum of 800 to 900 mbar e g Vacuum Pump cat no 84010 US and Canada 84000 Japan or 84020 rest of world are required for the protocol A Vacuum Regulator e g cat no 19530 should be used for easy monitoring of vacuum pressures and convenient vacuum release Removal of residual contaminants Nucleic acids remain bound to the membrane while contaminants are efficiently washed away during a sequence of wash steps In a single step highly pure viral RNA and DNA are eluted in Buffer AVE equilibrated to room temperature Elution of pure nucleic acids Elution is performed using Buffer AVE GlAamp MinElute columns allow minimal elution vol umes of only 20 pl Low elution volume leads to highly concentrated nucleic acid eluates For downstream applications that require small starting volumes e g some PCR and RT PCR assays a more concentrated eluate may increase assay sensitivity For downstream applications that require a larger starting volume the elution volume can be increased up to 150 pl However an increase in elution volume will decrease the concentration of nucleic acids in the eluate The eluate volume recovered can be up to 5 pl less than the volume of elution buffer applied to the column for example an elution buffer volume of 20 pl results in gt 15 pl final eluate The volume of eluate recovered depends on the nature of the sample Eluted DNA is collected in
8. 2931 proteins using QIAGEN spin column kits 1 year warranty on parts and labor For 48 viral nucleic acid preps 62724 Prefilled Reagent Cartridges Disposable Tip Holders Disposable Filter Tips Sample Tubes Elution Tubes Buffers Carrier RNA Preprogrammed card for EZ1 DSP 9017707 Virus protocol For 12 x 96 preps 12 QlAamp 965652 96 Plates RNase free Buffers QIAGEN Protease Elution Microtubes CL Caps S Blocks Carrier RNA For 12 x 96 preps 12 QlAamp 61762 96 Plates RNase Free Buffers QIAGEN Protease Elution Microtubes CL Caps S Blocks Carrier RNA Agreements for comprehensive service coverage are available please inquire QlAamp MinElute Virus Vacuum Handbook 04 2010 33 Ordering Information Product Contents Cat no GlAamp MinElute Media Kit 50 GlAamp Media MDx Kit 12 For 50 minipreps 50 QlAamp 57414 MinElute Columns QIAGEN Proteinase K Carrier RNA Buffers Extension Tubes 3 ml Collection Tubes 1 5 ml For 12 x 96 preps 12 GlAamp 259492 96 Plates Buffers Proteinase K S Blocks Disposable Troughs Racks with Elution Microtubes CL 0 4 ml Carrier RNA Top Elute Fluid Caps Tape Pad 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 giagen com or can be requested from QIAGEN Technical Services or your local distri
9. 500 pl of plasma or serum using the GlAamp MinElute Virus Vacuum Kit Important point before starting All centrifugation steps are carried out at room temperature 15 25 C Things to do before starting Equilibrate samples to room temperature Equilibrate Buffer AVE to room temperature for elution in step 20 Prepare a 56 C heating block for use in steps 4 and 19 Ensure that Buffer AW1 Buffer AW2 and QIAGEN Protease have been prepared according to instructions on pages 19 21 Add carrier RNA reconstituted in Buffer AVE to Buffer AL according to instructions on page 20 For processing using VacConnectors and VacValves set up the QlAvac 24 Plus as described on pages 16 18 Procedure Smp Pipet 75 pl QIAGEN Protease into a 2 ml microcentrifuge tube not provided Note Read Preparation of QIAGEN Protease page 20 for information about resuspending QIAGEN Protease in Buffer AVE recommended or Protease Resuspension Buffer Add 500 pl of plasma or serum into the 2 ml microcentrifuge tube Add 500 pl of Buffer AL containing 11 2 pg ml of Carrier RNA Close the cap and mix by pulse vortexing for 15 s In order to ensure efficient lysis it is essential that the sample and Buffer AL are mixed thoroughly to yield a homogeneous solution Note Do not add QIAGEN Protease directly to Buffer AL Incubate at 56 C for 15 min Briefly centrifuge the 2 ml tube to remove drops from the inside of the lid Add 600 pl of
10. QlAvac 24 Plus clean and dry For cleaning procedures see the QlAvac 24 Plus Handbook The components of the QlAvac 24 Plus are not resistant to certain solvents Table 1 If these solvents are spilled on the unit rinse it thoroughly with water To ensure consistent performance do not apply silicone or vacuum grease fo any part of the QlAvac 24 Plus manifold Always use caution and wear safety glasses when working near a vacuum manifold under pressure Contact QIAGEN Technical Services or your local distributor for information concerning spare or replacement parts The vacuum pressure is the pressure differential between the inside of the vacuum manifold and the atmosphere standard atmospheric pressure 1013 millibar or 760 mm Hg and can be measured using the QlAvac Connecting System or a vacuum regulator see Figure 1 The vacuum protocol requires a vacuum pump capable of producing a vacuum or 800 to 900 mbar e g QIAGEN Vacuum Pump Higher vacuum pressures must be avoided Use of vacuum pressures lower than recommended may reduce DNA yield and purity and increase the frequency of clogged membranes Table 1 Chemical resistance properties of QlAvac 24 Plus Resistant to Acetic acid Chaotropic salts Chlorine bleach Chromic acid Concentrated alcohols Hydrochloric acid SDS Sodium chloride Sodium hydroxide Tween 20 Urea Not resistant to Benzene Chloroform Ethers Phenol Toluene QlAamp MinElute Virus
11. Third Edition April 2010 GlAamp MinElute Virus Vacuum Handbook For simultaneous purification of viral RNA and DNA from plasma serum and cell free body fluids QIAGEN Trademarks QIAGEN GlAamp QlAcube BioRobot EZ1 MinElute QIAGEN Group Corex Corning Inc Tween ICI Americas Inc 2002 2007 QIAGEN all rights reserved QIAGEN is a member of the Forest Stewardship Council FSC For the production of printed materials eee including handbooks QIAGEN has a policy to select suppliers that comply with FSC standards for printing tne ae nado conc processes and well managed forests Contents Kit Contents 4 Storage 4 Quality Control 5 Product Use Limitations 5 Product Warranty and Satisfaction Guarantee 5 Technical Assistance 6 Safety Information 7 Introduction 8 Principle and procedure 8 Yield and size of viral nucleic acids 10 Carrier RNA 10 Addition of internal controls 12 Equipment and Reagents to Be Supplied by User 13 Important Notes 14 Handling of GlAamp MinElute columns 14 Centrifugation 19 Processing GlAamp MinElute columns in a microcentrifuge 19 Preparation of RNA 19 Sample storage 19 Preparation of QIAGEN Protease 20 Addition of carrier RNA to Buffer AL 20 Buffer AW 1 22 Buffer AW2 22 Elution of viral nucleic acids 22 Protocol E Purification of Viral Nucleic Acids from Plasma or Serum 23 Troubleshooting Guide 26 Appendix 30 Ordering Information 32 QlAamp MinElute Vi
12. This will reduce the failure rate of this type of tube during centrifugation Electrophoresis tanks Electrophoresis tanks should be cleaned with detergent solution e g 0 5 SDS rinsed with water dried with ethanol and then filled with a solution of 3 hydrogen peroxide After 10 minutes at room temperature the electrophoresis tanks should be rinsed thoroughly with RNase free water Solutions Solutions water and other solutions should be treated with 0 1 DEPC DEPC will react with primary amines and cannot be used directly to treat Tris buffers DEPC is highly unstable in the presence of Tris buffers and decomposes rapidly into ethanol and CO When preparing Tris buffers treat water with DEPC first and then dissolve Tris to make the appropriate buffer DEPC is a strong but not absolute inhibitor of RNases It is commonly used at a concentration of 0 1 to inactivate RNases on glass or plasticware or to create RNase free solutions and water DEPC inactivates RNases by covalent modification Trace amounts of DEPC will modify purine residues in RNA by carbethoxylation Carbethyoxylated RNA is translated with very low efficiency in cell free systems However its ability to form DNA RNA or RNA RNA hybrids is not seriously affected unless a large fraction of the purine residues have been modified Residual DEPC must always be removed from solutions or vessels by autoclaving or heating to 100 C for 15 minutes Add 0 1 ml DEPC to 1
13. Vacuum Handbook 04 2010 15 Regulator gauge Knob Figure 1 Schematic diagram of the Vacuum Regulator Setup of the AlAvac 24 Plus vacuum manifold 1 Connect the QlAvac 24 Plus to a vacuum source If using the QlAvac Connecting System connect the system to the manifold and vacuum source as described in Appendix A of the QlAvac 24 Plus Handbook 2 Insert a VacValve into each luer slot of the QlAvac 24 Plus that is to be used see Figure 2 Close unused luer slots with luer plugs or close the inserted VacValve VacValves should be used if flow rates of samples differ significantly to ensure consistent vacuum 16 QlAamp MinElute Virus Vacuum Handbook 04 2010 Insert a VacConnector into each VacValve see Figure 2 Perform this step directly before starting the purification to avoid exposure of VacConnectors to potential contaminants in the air Place the GlAamp MinElute columns into the VacConnectors on the manifold see Figure 2 Insert an Extension Tube into each AlAamp MinElute column see Figure 3 For nucleic acid purification follow the instructions in the vacuum protocol Discard the VacConnectors appropriately after use Leave the lid of the QlAamp MinElute column open while applying vacuum Switch off the vacuum between steps to ensure that a consistent even vacuum is applied during processing For faster vacuum release a vacuum regulator should be used see Figure 1 Note Each VacValve can be c
14. and do not require pretreatment to inactivate RNases Nondisposable plasticware Nondisposable plasticware should be treated before use to ensure that it is RNase free Plasticware should be thoroughly rinsed with 0 1 M NaOH 1 mM EDTA followed by RNase free water see Solutions page 31 Alternatively chloroform resistant plasticware can be rinsed with chloroform to inactivate RNases Glassware Glassware should be treated before use to ensure that it is RNase free Glassware used for RNA work should be cleaned with detergent thoroughly rinsed and oven baked at gt 240 C for four or more hours overnight if more convenient before use Autoclaving alone will not fully inactivate many RNases Oven baking will both inactivate ribonucleases and ensure that no other nucleic acids such as plasmid DNA remain on the surface When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information consult the appropriate material safety data sheets MSDSs available from the product supplier 30 QlAamp MinElute Virus Vacuum Handbook 04 2010 of the glassware Alternatively glassware can be treated with DEPC diethyl pyrocar bonate Cover the glassware with 0 1 DEPC in water overnight 12 hours at 37 C and then autoclave or heat to 100 C for 15 minutes to remove residual DEPC Note Corex tubes should be rendered RNase free by treatment with DEPC and not by baking
15. ate Carefully open the lid of the QlAamp MinElute column and apply 20 150 pl of Buffer AVE or RNase free water to the center of the membrane Close the lid and incubate at room temperature for 1 min Centrifuge at full speed 20 000 x g 14 000 rpm for 1 min Important Ensure that the elution buffer is equilibrated to room temperature If elution is done in small volumes lt 50 pl the elution buffer must be dispensed onto the center of the membrane for complete elution of bound RNA and DNA Elution volume is flexible and can be adapted according to the requirements of the downstream applications Remember that the recovered eluate volume will differ by approximately 5 pl from the elution buffer volume applied onto the column Incubating the GlAamp MinElute column loaded with Buffer AVE or water for 5 min at room temperature before centrifugation generally increases DNA and RNA yield QlAamp MinElute Virus Vacuum Handbook 04 2010 25 JO20jO1g Troubleshooting Guide This troubleshooting guide may be helpful in solving any problems that may arise The scientists in QIAGEN Technical Services are always happy to answer any questions you may have about either the information and protocol in this handbook or molecular biology applications see back cover for contact information Comments and suggestions Little or no nucleic acid in the eluate a Carrier RNA not added to Buffer AL b Degraded carrier RNA c Buffer AL carrie
16. but only until the kit expiration date Keeping the GIAGEN Protease stock solution at room temperature for prolonged periods of time should be avoided Storage at 20 C will prolong its life but repeated freezing and thawing should be avoided Dividing the solution into aliquots and freezing at 20 C is recommended Label the aliquots and indicate which buffer was used for resuspension Addition of carrier RNA to Buffer AL Add 310 pl Buffer AVE to the tube containing 310 pg lyophilized carrier RNA to obtain a solution of 1 pg pl Dissolve the carrier RNA thoroughly divide it into conveniently sized aliquots and store it at 20 C Do not freeze thaw the aliquots of carrier RNA more than 3 times Note that carrier RNA does not dissolve in Buffer AL It must first be dissolved in Buffer AVE and then added to Buffer AL Calculate the volume of Buffer AL carrier RNA mix needed per batch of samples by selecting the number of samples to be simultaneously processed from Table 2 For larger numbers of samples volumes can be calculated using the following sample calculation Contains chaotropic salt Take appropriate laboratory safety measures and wear gloves when handling Not compatible with disinfectants containing bleach See page 7 for safety information 20 QlAamp MinElute Virus Vacuum Handbook 04 2010 n x 0 55 ml y ml y ml x 11 2 pl ml z pl where n number of samples to be processed simultaneously y calculated volume of Buff
17. butor 34 QlAamp MinElute Virus Vacuum Handbook 04 2010 Notes QlAamp MinElute Virus Vacuum Handbook 04 2010 35 www giagen 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 2 1 3865 3865 Fax 86 21 3865 3965 Technical 800 988 0325 Denmark Orders 80 885945 Fax 80 885944 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 1 122 330 Technical 01 800 7742 639 The Netherlands
18. e Repeated freezing and thawing should be avoided see page 19 Always use fresh samples or samples thawed only once Samples were left standing at room temperature for too long Repeat the purification procedure with new samples Reconstituted QIAGEN Protease was subjected to elevated temperature for a prolonged time Repeat the procedure using new samples and fresh QIAGEN Protease QlAamp MinElute Virus Vacuum Handbook 04 2010 27 Comments and suggestions eJ Too much or too little carrier RNA in the eluate f Reduced sensitivity g Performance of purified nucleic acids in downstream assays varies with aging of reconstituted wash buffers h Anew combination of reverse transcriptase and Tay DNA polymerase was used General handling a Clogged GlAamp MinElute column b Variable elution volumes 28 Determine the maximum amount of carrier RNA suitable for your amplification reaction Adjust the concentration of carrier RNA added to Buffer AL accordingly see Addition of carrier RNA to Buffer AL page 20 Determine the maximum volume of eluate suitable for your amplification reaction Reduce or increase the volume of eluate added to the amplification reaction accordingly The elution volume can be adapted proportionally Salt and ethanol components of Buffers AW1 and AW2 may have separated out after being left for a long period between preparations Always mix buffers thoroughly before each preparat
19. er AL z volume of carrier RNA Buffer AVE to add to Buffer AL Gently mix by inverting the tube 10 times To avoid foaming do not vortex Table 2 Volumes of Buffer AL and carrier RNA Buffer AVE mix required for the QlAamp MinElute Virus Vacuum procedure No Vol Buffer Vol Carrier No Vol Buffer Vol Carrier samples AL ml RNA AVE pl samples AL ml RNA AVE pl 0 55 6 2 13 7 495 80 1 2 1 10 12 3 14 7 70 86 2 3 1 65 18 5 15 8 25 92 4 4 2 20 24 6 16 8 80 98 6 5 2 75 30 8 17 9 35 104 7 6 3 30 37 0 18 9 90 110 9 7 3 85 43 1 19 10 45 117 0 8 4 40 49 3 20 11 00 1232 9 4 95 55 0 21 11 55 129 4 10 5 50 61 6 22 1220110 135 5 11 6 05 67 8 23 12 65 141 7 12 6 60 73 9 24 13 20 147 8 Note The sample preparation procedure is optimized for 5 6 pg of carrier RNA per sample If less carrier RNA has been shown to be better for your amplification system transfer only the required amount of dissolved carrier RNA to the tubes containing Buffer AL For each microgram of carrier RNA required per preparation add 2 yl Buffer AVE dissolved carrier RNA per milliliter of Buffer AL Use of less than 5 6 pg carrier RNA per sample must be validated for each particular sample type and downstream assay QlAamp MinElute Virus Vacuum Handbook 04 2010 21 Buffer AW1 Add 25 ml of ethanol 96 100 to a bottle containing 19 ml of Buffer AW concentrate as described on the bottle Tick the check box on the label to indicate that
20. ethanol 96 100 to the sample Close the cap and mix thoroughly by pulse vortexing for 15 s Incubate the lysate with the ethanol for 5 min at room temperature 15 25 C Note If ambeint temperature exceeds 25 C ethanol should be cooled on ice before adding to the lysate QlAamp MinElute Virus Vacuum Handbook 04 2010 23 JO20jO1g o 2 o a 10 11 12 13 14 24 Briefly centrifuge the 2 ml tube to remove drops from the inside of the lid Insert the AlAamp MinElute column into the VacConnector on the AlAvac 24 Plus Insert an extension tube into the open QlAamp MinElute column Note Keep the collection tube for the dry spin in step 18 Make sure that the main vacuum valve between the vacuum pump and the vacuum manifold and the screw cap valve on the end of the QlAvac 24 Plus vacuum manifold are closed Switch on the vacuum pump by pressing the power switch The vacuum is applied only to the connecting system if used and not to the vacuum manifold Note For fast and convenient release of the vacuum pressure the QlAvac Connect ing System or the Vacuum Regulator should be used see Ordering Information page 32 Carefully apply all of the lysate from step 7 into the extension tube of the QlAamp MinElute column without wetting the rim Avoid touching the QlAamp MinElute column membrane with the pipet tip Open the main vacuum valve After all lysates have been drawn through the GlAa
21. ethanol has been added Store reconstituted Buffer AW at room temperature 15 25 C Reconstituted Buffer AW is stable for up to 1 year when stored at room temperature but only until the kit expiration date Note Always mix reconstituted Buffer AW 1 by shaking before starting the procedure Buffer AW2 Add 30 ml of ethanol 96 100 to a bottle containing 13 ml of Buffer AW2 concentrate as described on the bottle Tick the check box on the label to indicate that ethanol has been added Store reconstituted Buffer AW2 at room temperature 15 25 C Reconstituted Buffer AW2 is stable for up to 1 year when stored at room temperature but only until the kit expiration date Note Always mix reconstituted Buffer AW2 by shaking before starting the procedure Elution of viral nucleic acids Elution buffer should be equilibrated to room temperature before it is applied to the column Yields will be increased if the QlAamp MinElute column is incubated with the elution buffer at room temperature for 5 minutes before centrifugation Contains chaotropic salt Take appropriate laboratory safety measures and wear gloves when handling Not compatible with disinfectants containing bleach See page 7 for safety information t Contains sodium azide as a preservative 22 QlAamp MinElute Virus Vacuum Handbook 04 2010 Protocol Purification of Viral Nucleic Acids from Plasma or Serum This protocol is for purification of viral nucleic acids from
22. g the GlAamp MinElute Virus Vacuum protocols in combination with commercially available amplification systems may require the introduction of an internal control into the purification procedure Internal control RNA or DNA should be added together with the carrier RNA to the lysis buffer For optimal purification efficiency internal control molecules should be longer than 200 nucleotides as smaller molecules are not efficiently recovered Refer to the manufacturer s instructions in order to determine the optimal concentration Using a concentration other than that recommended may reduce amplification efficiency 12 QlAamp MinElute Virus Vacuum Handbook 04 2010 Eguipment and Reagents to Be Supplied by User When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information consult the appropriate material safety data sheets MSDSs available from the product supplier Ethanol 96 100 2 ml microcentrifuge tubes Pipets and pipet tips pipet tips with aerosol barriers for preventing cross contamination are recommended Heating block for lysis of samples at 56 C Microcentrifuge with rotor for 1 5 ml and 2 ml tubes Vortexer QlAvac 24 Plus vacuum manifold cat no 19413 or equivalent VacConnectors cat no 19407 Vacuum Regulator cat 19530 for easy monitoring of vacuum pressures and easy releasing of vacuum Vacuum Pump cat no 84010 USA and Canada 84000 Japa
23. hnical assistance and more information please call one of the QIAGEN Technical Service Departments or local distributors see back cover 6 QlAamp MinElute Virus Vacuum Handbook 04 2010 Safety Information 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 ts msds asp 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 waste containing Buffer AL or Buffer AW1 Buffers AL and AW contain guanidine hydrochloride 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 the GlAamp MinElute Virus Vacuum Kit Buffers AL and AW1 Contains guanidine hydrochloride harmful irritant Risk and safety phrases R22 36 38 513 26 36 46 QIAGEN Protease Contains subtilisin sensitizer irritant Risk and safety phrases R37 38 4 1 42 522 24 26 36 37 39 46 24 hour emergency informatio
24. ication of high quality viral nucleic acids The GlAcube is preinstalled with protocols for purification of plasmid DNA genomic DNA RNA viral nucleic acids and proteins plus DNA and RNA cleanup The range of protocols available is continually expanding and additional QIAGEN protocols can be downloaded free of charge at www giagen com MyGlAcube Sample volumes using the QlAamp MinElute Virus Vacuum Kit Each GlAamp MinElute column can bind nucleic acids that are longer than 200 bases but yield depends on sample volume and virus titer The vacuum procedure is optimized for use with a starting volume of 500 yl 8 QlAamp MinElute Virus Vacuum Handbook 04 2010 Lysis with QIAGEN Protease Samples are lysed under highly denaturing conditions at elevated temperatures Lysis is performed in the presence of QIAGEN Protease and Buffer AL which together ensure inactivation of RNases Adsorption to the AlAamp MinElute membrane Binding conditions are adjusted by adding ethanol to allow optimal binding of the viral RNA and DNA to the membrane Lysates are then transferred onto a QlAamp MinElute column and viral nucleic acids are adsorbed onto the silica gel membrane as the lysate is drawn through by vacuum pressure Salt and pH conditions ensure that protein and other contaminants which can inhibit PCR and other downstream enzymatic reactions are not retained on the QlAamp MinElute membrane A vacuum manifold e g QlAvac 24 Plus cat no
25. ion If enzymes are changed it may be necessary to readjust the amount of carrier RNA added to Buffer AL and the amount of eluate used Remove the GlAamp MinElute column from the vacuum manifold place it in a 2 ml collection tube and centrifuge it at full speed until sample has completely passed the membrane Cryoprecipitates may have formed in plasma due to repeated freezing and thawing These can block the QlAamp MinElute column Do not use plasma that has been frozen and thawed more than once In case cryoprecipitates are visible clear the sample by centrifugation as described in Sample storage on page 19 before starting the sample preparation This is normal when different sample types have been processed QlAamp MinElute Virus Vacuum Handbook 04 2010 Comments and suggestions c Vacuum pressure of 800 900 mbar not reached Gasket of QlAvac lid has worn out Check the seal of the manifold visually and replace it if necessary VacValves have worn out Remove all VacValves and insert VacConnectors directly into the luer extensions Insert QlAamp MinElute columns into VacConnectors close the lid of the columns and switch on vacuum Check if vacuum pressure is reached Replace VacValves if necessary Connection to vacuum pump is leaky Close all luer extension with luer caps and switch on the vacuum pump Check if vacuum pressure is stable after the pump is switched on and the Vacuum Regulator valve
26. is closed Exchange the connections between pump and vacuum manifold if necessary After all above checks have been made replace the vacuum pump with a stronger one QlAamp MinElute Virus Vacuum Handbook 04 2010 29 Appendix Handling RNA Ribonucleases RNases are very stable and active enzymes that generally do not reguire 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 Care should be taken to avoid inadvertently introducing RNases into the RNA sample during or after the isolation procedure In order to create and maintain an RNase free environment the following precautions must be taken during pretreatment and use of disposable and non disposable vessels and solutions while working with RNA General handling Proper microbiological aseptic technique should always be used when working with RNA Hands and dust particles may carry bacteria and molds and are the most common sources of RNase contamination Always wear latex or vinyl gloves while handling reagents and RNA samples to prevent RNase contamination from the surface of the skin or from dusty laboratory equipment Change gloves frequently and keep tubes closed Disposable plasticware The use of sterile disposable polypropylene tubes is recommended throughout the procedure These tubes are generally RNase free
27. losed individually when the sample is completely drawn through the spin column allowing parallel processing of samples of different volumes or viscosities After processing samples clean the QlAvac 24 Plus see Cleaning and Decontaminating the QlAvac 24 Plus in the AlAvac 24 Plus Handbook Note Buffers AL and AW1 used in GlAamp MinElute procedure are not compatible with disinfecting agents containing bleach See page 7 for safety information QlAamp MinElute Virus Vacuum Handbook 04 2010 17 Figure 2 Setting up the GlAvac 24 Plus with AlAamp MinElute columns using VacValves and VacConnectors 1 QlAvac 24 Plus vacuum manifold 4 VacConnector 2 Luer slot of the QlAvac 24 Plus 5 QlAamp column 3 VacValve 6 Luer slot closed with luer plug Must be purchased separately Figure 3 Assembly of components of the AlAamp MinElute Vacuum Kit 1 VacValvet 2 VacConnectort 2 p 1 3 GlAamp MinElute column 4 Extension tube Must be purchased separately 18 QlAamp MinElute Virus Vacuum Handbook 04 2010 Centrifugation For the dry spin at the end of the washing procedure and for elution centrifugation should be carried out at full speed All centrifugation steps should be carried out at room temperature 15 25 C Processing QlAamp MinElute columns in a microcentrifuge M Close the QlAamp MinElute column before placing it in the microcentrifuge Centrifuge as described M Remove the QlAamp Mi
28. mp MinElute column close the main vacuum valve and open the screw cap valve to vent the manifold Close the screw cap valve after the vacuum is released from the manifold After closing the main vacuum valve the vacuum is applied only to the connecting system if used and not the vacuum manifold If the lysates from individual samples have not completely passed through the membrane despite the VacValves of all other QlAamp MinElute columns being closed place the QlAamp MinElute column into a clean 2 ml collection tube not provided close the cap and centrifuge at full speed for 3 min or until it has completely passed through Additional collection tubes can be purchased separately see Ordering Information page 32 Apply 600 pl of Buffer AW1 to the QlAamp MinElute column without wetting the rim Avoid touching the AlAamp MinElute column membrane with the pipet tip Remove and discard the extension tube Note To avoid cross contamination take care not to take tubes across neighboring GlAamp MinElute columns during extension tube removal Open the main vacuum valve After all Buffer AW1 has been drawn through the QlAamp MinElute column close the main vacuum valve and open the screw cap valve to vent the manifold Close the screw cap valve after the vacuum is released from the manifold Apply 750 pl of Buffer AW2 to the AlAamp MinElute column without wetting the rim Avoid touching the GIAamp MinElute column membrane with the pi
29. n Emergency medical information in English French and German can be obtained 24 hours a day from Poison Information Center Mainz Germany Tel 49 6131 19240 R22 Harmful if swallowed R36 38 Irritating to eyes and skin R37 38 Irritating to respiratory system and skin R41 Risk of serious damage to eyes R42 May cause sensitization by inhalation 13 Keep away from food drink and animal feeding stuffs S22 Do not breathe dust S24 Avoid contact with skin S26 In case of contact with eyes rinse immediately with plenty of water and seek medical advice S36 Wear suitable protective clothing S36 37 39 Wear suitable protective clothing gloves and eye face protection S46 If swallowed seek medical advice immediately and show the container or label QlAamp MinElute Virus Vacuum Handbook 04 2010 7 Introduction The GlAamp MinElute Virus Vacuum Kit uses well established technology for simultaneous purification of viral DNA and RNA The kit combines the selective binding properties of a silicatbased membrane with flexible elution volumes of between 20 and 150 pl The procedure is suitable for use with plasma serum or other cell free body fluids Samples can be either fresh or frozen provided they have not been frozen and thawed more than once see page 19 Viral nucleic acids are eluted in Buffer AVE ready for use in amplification reactions or storage at 20 C Purified nucleic acids are free of proteins nucleases and othe
30. n or 84020 rest of world or equivalent pump capable of producing a vacuum of 800 to 900 mbar Optional VacValves cat no 19408 Optional QlAvac Connecting System cat no 19419 Do not use denatured alcohol which contains other substances such as methanol or methylethylketone QlAamp MinElute Virus Vacuum Handbook 04 2010 13 Important Notes Handling of AlAamp MinElute columns Because of the sensitivity of nucleic acid amplification technologies the following precautions are necessary when handling QlAamp MinElute columns in order to avoid cross contamination between sample preparations E Carefully apply the sample or solution to the QlAamp MinElute column Pipet the sample into the GlAamp MinElute column without wetting the rim of the column E Change pipet tips between all liquid transfers The use of aerosol barrier pipet tips is recommended E Avoid touching the GlAamp MinElute membrane with the pipet tip E After all pulse vortexing steps briefly centrifuge the microcentrifuge tubes to remove drops from the inside of the lid E Wear gloves throughout the entire procedure In case of contact between gloves and sample change gloves immediately The QlAvac 24 Plus The QlAvac 24 Plus is designed for fast and efficient vacuum processing of up to 24 QIAGEN spin columns in parallel Samples and wash solutions are drawn through the column membranes by vacuum instead of centrifugation providing greater speed and
31. nElute column and collection tube from the microcentrifuge Discard the filtrate and the used 2 ml collection tube Place the QlAamp MinElute column in the 1 5 ml collection tube B Open only one GlAamp MinElute column at a time and take care to avoid generating aerosols Preparation of RNA When preparing viral RNA work quickly during the manual steps of the procedure If you have not previously worked with RNA read the Appendix on page 30 before starting Buffer AVE is RNase free upon delivery It contains sodium azide an antimicrobial agent that prevents growth of RNase producing organisms However as this buffer does not contain any RNase inhibitors it will not actively inhibit RNases introduced by inappropriate handling Extreme care should be taken to avoid contamination with RNases when handling Buffer AVE Sample storage After collection and centrifugation plasma or serum can be stored at 2 8 C for up to 6 hours For long term storage freezing at 20 C or 80 C in aliquots is recommended Frozen plasma or serum samples must not be thawed more than once Repeated freeze thawing leads to denaturation and precipitation of proteins resulting in reduced viral titers and therefore reduced yields of viral nucleic acids In addition cryoprecipitates formed during freeze thawing will clog the QlAamp MinElute membrane If cryoprecipitates are visible they can be pelleted by centrifugation at 6800 x g for 3 minutes The cleared supe
32. ormance QIAGEN Protease reconstituted in Buffer AVE or Protease Resuspension Buffer is stable for up to 1 year when stored at 2 8 C but only until the kit expiration date Keeping the QIAGEN Protease stock solution at room temperature for prolonged periods of time should be avoided 4 QlAamp MinElute Virus Vacuum Handbook 04 2010 Quality Control In accordance with GIAGEN s ISO certified Quality Management System each lot of QlAamp MinElute Virus Vacuum Kits is tested against predetermined specifications to ensure consistent product quality Product Use Limitations The GlAamp MinElute Virus Vacuum Kit is intended for molecular biology applications This product is not intended for the diagnosis prevention or treatment of a disease All due care and attention should be exercised in the handling of the products We recommend all users of QIAGEN products to adhere to the NIH guidelines that have been developed for recombinant DNA experiments or to other applicable guidelines Product Warranty and Satisfaction Guarantee QIAGEN guarantees the performance of all products in the manner described in our product literature The purchaser must determine the suitability of the product for its particular use Should any product fail to perform satisfactorily due to any reason other than misuse QIAGEN will replace it free of charge or refund the purchase price We reserve the right to change alter or modify any product to enhance its perf
33. ormance and design If a QIAGEN product does not meet your expectations simply call your local Technical Service Department or distributor We will credit your account or exchange the product as you wish Separate conditions apply to QIAGEN scientific instruments service products and to products shipped on dry ice Please inquire for more information A copy of QIAGEN terms and conditions can be obtained on request and is also provided on the back of our invoices If you have questions about product specifications or performance please call QIAGEN Technical Services or your local distributor see back cover QlAamp MinElute Virus Vacuum Handbook 04 2010 5 Technical Assistance At GIAGEN we pride ourselves on the guality and availability of our technical support Our Technical Service Departments are staffed by experienced scientists with extensive practical and theoretical expertise in molecular biology and the use of QIAGEN products If you have any questions or experience any difficulties regarding the GlAamp MinElute Virus Vacuum 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 performance or new applications and techniques For tec
34. pet tip Leave the lid of the column open QlAamp MinElute Virus Vacuum Handbook 04 2010 20 Open the main vacuum valve After all Buffer AW2 has been drawn through the QlAamp MinElute column close the main vacuum valve and open the screw cap valve to vent the manifold Close the screw cap valve after the vacuum is released from the manifold Apply 750 pl of ethanol 96 100 to the AlAamp MinElute column without wetting the rim Avoid touching the AlAamp MinElute column membrane with the pipet tip Leave the lid of the column open Open the main vacuum valve After all ethanol has been drawn through the QlAamp MinElute column close the main vacuum valve and open the screw cap valve to vent the manifold Close the screw cap valve after the vacuum is released from the manifold Close the lid of the AlAamp MinElute column Remove it from the vacuum manifold and discard the VacConnector Place the QlAamp MinElute column in a clean 2 ml collection tube saved from step 8 and centrifuge at full speed 20 000 x g 14 000 rpm for 3 min to dry the membrane completely Recommended Place the QlAamp MinElute column into a new 2 ml collection tube not provided open the lid and incubate the assembly at 56 C for 3 min to dry the membrane completely This step serves to evaporate any remaining liguid Place the QlAamp MinElute column in a clean 1 5 ml microcentrifuge tube provided and discard the collection tube with the filtr
35. r RNA mixture mixed insufficiently d Low percentage ethanol used instead of 96 100 e RNA degraded 26 Reconstitute carrier RNA in Buffer AVE and mix with Buffer AL as described on page 20 Repeat the purification procedure with new samples Carrier RNA reconstituted in Buffer AVE was not stored at 20 C or underwent multiple freeze thaw cycles Alternatively Buffer Al carrier RNA mixture was stored for more than 48 hours at 2 8 C Prepare a new tube of carrier RNA dissolved in Buffer AVE and mix with Buffer AL Repeat the purification procedure with new samples Mix Buffer AL with carrier RNA by gently inverting the tube of Buffer AL carrier RNA at least 10 times Repeat the purification procedure with new samples and 96 100 ethanol Do not use denatured alcohol which contains other substances such as methanol or methylethylketone Check the integrity of the RNA in the original samples Often RNA is degraded by RNases in the starting material plasma serum body fluids Ensure that the samples are processed quickly following collection or removal from storage Check for RNase contamination of buffers and water and ensure that no RNase is introduced during the procedure Use Buffer AVE or RNase free water for elution QlAamp MinElute Virus Vacuum Handbook 04 2010 Comments and suggestions f RNase contamination be careful to in Buffer AVE g Buffer AW1 or AW2 prepared incorrectly h Buffer AW1 or
36. r impurities Principle and procedure The GlAamp MinElute Virus Vacuum procedure comprises 4 steps lyse bind wash elute and is carried out using GlAamp MinElute columns on a vacuum manifold The procedure is designed to ensure that there is no sample to sample cross contamination and allows safe handling of potentially infectious samples The simple GlAamp MinElute Vacuum procedure which is highly suited for simultaneous processing of multiple samples yields pure nucleic acid in less than 1 hour The AlAamp MinElute Virus Vacuum Kit can be used for isolation of viral RNA and DNA from a broad range of RNA and DNA viruses However performance cannot be guaranteed for every virus species and must be validated by the customer Automated viral nucleic purification on the QlAcube using the AlAamp MinElute Virus Spin Kit Purification of viral nucleic acids using the AlAamp MinElute procedure can now be fully automated on the new GlAcube using the QlAamp MinElute Virus Spin Kit cat no 57704 The kit uses the same chemistries as the GlAamp MinElute Virus Vacuum Kit but with smaller sample volumes The innovative QlAcube uses advanced technology to process QIAGEN spin columns enabling seamless integration of automated low throughput sample prep into your laboratory workflow The GlAcube performs the same steps as the manual spin procedure lyse bind wash and elute enabling you to continue using GlAamp MinElute Virus chemistries for purif
37. rnatant should be removed and processed immediately without disturbing the pellet This step will not reduce viral titers QlAamp MinElute Virus Vacuum Handbook 04 2010 19 Preparation of QIAGEN Protease This kit provides two alternative buffers for dissolving QIAGEN Protease Buffer AVE recommended or Protease Resuspension Buffer Dissolving the protease in Buffer AVE provides a generic and efficient working solution for all starting materials As an alternative dissolving QIAGEN Protease in Protease Resuspension Buffer provides efficient viral lysis for most sample types For some starting materials such as EDTA plasma performance is slightly enhanced However Protease Resuspension Buffer is not compatible with samples or internal controls that contain phosphate e g viral transport medium cell culture supernatants or phosphate buffered saline If the sample or internal control contains phosphate it is highly recommended to resuspend QIAGEN Protease in Buffer AVE Add 4 4 ml of Buffer AVE or Protease Resuspension Buffer to the vial of lyophilized QIAGEN Protease and mix carefully to avoid foaming Make sure that the QIAGEN Protease is completely dissolved Label the resuspended QIAGEN Protease to indicate which buffer was used for resuspension Note Do not add QIAGEN Protease directly to Buffer AL QIAGEN Protease reconstituted in Buffer AVE or Protease Resuspension Buffer is stable for 12 months when stored at 2 8 C
38. rus Vacuum Handbook 04 2010 3 Kit Contents QlAamp MinElute Virus Vacuum Kit 50 Catalog no 57714 Number of preps 50 GlAamp MinElute Columns 50 Extension Tubes 3 ml 50 Collection Tubes 1 5 ml 50 Buffer AL 33 ml Buffer AW1 concentrate 19 ml Buffer AW2 concentrate 13 ml Buffer AVE tubes with purple caps 7 x2 ml Protease Resuspension Buffer 6 ml Carrier RNA tubes with red caps 310 yg QIAGEN Protease 1 vial Handbook 1 Contains a chaotropic salt Take appropriate laboratory safety measures and wear gloves when handling Not compatible with disinfectants containing bleach See page 7 for safety information t Contains sodium azide as a preservative Resuspension volume 4 4 ml See Preparation of QIAGEN Protease page 20 Storage QlAamp MinElute columns should be stored at 2 8 C upon arrival All buffers can be stored at room temperature 15 25 C Lyophilized carrier RNA can be stored at room temperature 15 25 C until the expiration date on the kit box Carrier RNA can only be dissolved in Buffer AVE dissolved carrier RNA should be immediately added to Buffer AL as described on page 20 This solution should be prepared fresh and is stable at 2 8 C for up to 48 hours Unused portions of carrier RNA dissolved in Buffer AVE should be frozen in aliquots at 20 C Lyophilized QIAGEN Protease can be stored at room temperature 15 25 C until the kit expiration date without affecting perf
39. vac Connecting System System to connect vacuum manifold 19419 with vacuum pump includes Tray Waste Bottles Tubing Couplings Valve Gauge 24 VacValves Vacuum Pump Universal vacuum pump Inquire Vacuum Regulator For use with GlAvac manifolds 19530 GIAGEN Protease 7 5 AU 7 5 Anson Units lyophilized VAS QIAGEN Protease 30 AU 4x7 5 Anson Units lyophilized 19157 Buffer AL 216 ml 216 ml Lysis Buffer AL 19075 Buffer AW 1 concentrate 242 ml 242 ml Wash Buffer 1 Concentrate 19081 Buffer AW2 concentrate 324 ml 324 ml Wash Buffer 2 Concentrate 19072 Collection Tubes 2 ml 1000 collection tubes 2 ml 19201 Extension Tubes 3 ml For use with QIAGEN Mini or 19587 32 MinElute columns on vacuum manifold 100 per pack QlAamp MinElute Virus Vacuum Handbook 04 2010 Ordering Information Product Contents Cat no Related products QlAamp DSP Virus Kit For 50 minipreps QlAamp 60704 QlAamp MinElute Virus Spin Kit 50 QlAcube 110 V QlAcube 230 V EZ1 DSP Virus Kit 48 EZ1 DSP Virus Card QlAamp Virus BioRobot MDx Kit 12 QlAamp DSP 96 Virus MDx Kit US Canada and Japan t Rest of world MinElute Columns Buffers Reagents Tubes Column Extenders VacConnectors For 50 manual or fully automated 57704 minipreps 50 GlAamp MinElute Columns QIAGEN Protease Carrier RNA Buffers Collection Tubes 2 ml Robotic workstation for automated 9001292 purification of nucleic acids or 9001
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