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  • QIAamp DNA Blood Midi/MaxiHandbook

    For large-scale genomic and viral DNA purificationfrom

    whole bloodplasmaserumbody fluidslymphocytes

    Third Edition April 2010

    W W W . Q I A G E N . C O M

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  • Trademarks: QIAGEN, QIAamp, BioRobot, RNeasy (QIAGEN Group); Combitips, Eppendorf, Multipette (Eppendorf-Netheler-Hinz GmbH); Seripettor (Brand GmbH and Co KG).

    QIAGEN robotic systems are not available in all countries; please inquire.

    The PCR process is covered by U. S. Patents 4,683,195 and 4,683,202 and foreign equivalents owned by Hoffmann-La Roche AG.

    20002010 QIAGEN, all rights reserved.

    QIAGEN is a member of the Forest Stewardship Council (FSC). For the production of printed materials,including handbooks, QIAGEN has a policy to select suppliers that comply with FSC standards for printingprocesses and well-managed forests.

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  • QIAamp DNA Blood Midi/Maxi Handbook 04/2010 3

    ContentsKit Contents 4

    Storage 5

    Product Use Limitations 5

    Product Warranty and Satisfaction Guarantee 5

    Technical Assistance 6

    Quality Control 6

    Safety Information 7

    Introduction 8

    The QIAamp Midi/Maxi procedure 8

    Equipment and Reagents to Be Supplied by User 14

    Important Notes 15

    Centrifugation 15

    Processing QIAamp Midi and QIAamp Maxi spin columns on the QIAvac 24 Plus 15

    Preparation of reagents 18

    Optimized handling for higher throughputs 19

    Protocols

    Purification of DNA from Whole Blood Using the QIAamp DNA Blood Midi Kit (Spin Protocol) 20

    Purification of DNA from Whole Blood Using the QIAamp DNA Blood Maxi Kit (Spin Protocol) 24

    Purification of DNA from Whole Blood Using the QIAamp DNA Blood Midi Kit (Vacuum Protocol) 28

    Purification of DNA from Whole Blood Using the QIAamp DNA Blood Maxi Kit (Vacuum Protocol) 31

    Troubleshooting Guide 34

    Appendix: Determination of Concentration, Yield, Purity, and Length of DNA 38

    Ordering Information 39

    QIAGEN Distributors 43

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  • Kit Contents

    QIAamp DNA Blood Midi Kit (20) (100)

    Catalog no. 51183 51185

    Number of preps 20 100

    QIAamp Midi Spin Columns 20 100

    Collection Tubes (15 ml) 20 100

    Buffer AL* 54 ml 265 ml

    Buffer AW1* (concentrate) 19 ml 95 ml

    Buffer AW2 (concentrate) 17 ml 66 ml

    Buffer AE 12 ml 60 ml

    QIAGEN Protease 1 vial 4 vials

    Handbook 1 1

    QIAamp DNA Blood Maxi Kit (10) (50)

    Catalog no. 51192 51194

    Number of preps 10 50

    QIAamp Maxi Spin Columns 10 50

    Collection Tubes (50 ml) 10 50

    Buffer AL* 126 ml 2 x 330 ml

    Buffer AW1* (concentrate) 27 ml 121 ml

    Buffer AW2 (concentrate) 17 ml 81 ml

    Buffer AE 22 ml 110 ml

    QIAGEN Protease 1 vial 5 vials

    Handbook 1 1

    * Not compatible with disinfecting agents containing bleach. Contains a chaotropic salt. Take appropriate safetymeasures, and see page 7 for further safety information.

    Resuspension volume 4.4 ml Resuspension volume 5.5 ml

    QIAamp DNA Blood Midi/Maxi Handbook 04/20104

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  • QIAamp DNA Blood Midi/Maxi Handbook 04/2010 5

    Storage QIAamp Midi spin columns and buffers can be stored dry at room temperature(1525C). Upon arrival, store QIAamp Maxi spin columns at 28C. All otherQIAamp Maxi components can be stored at 28C or at room temperature (1525C).Kits can be stored under these conditions for up to 1 year without showing any reductionin performance. For longer periods, storing the QIAamp DNA Blood Midi and MaxiKits at 28C is recommended.

    Lyophilized QIAGEN Protease can be stored at room temperature (1525C) for up to 1 year without any reduction in performance. For storage longer than 1 year or if ambienttemperatures constantly exceed 25C, QIAGEN Protease should be stored dry at 28C.

    Reconstituted QIAGEN Protease is stable for 2 months when stored at 28C. Keepingthe QIAGEN Protease stock solution at room temperature for prolonged periods of timeshould be avoided. Storage at 20C will prolong its life, but repeated freezing andthawing should be avoided. Dividing the solution into usable aliquots and storage at 20Cis recommended.

    Product Use LimitationsQIAamp DNA Blood Kits are intended for molecular biology applications. These productsare 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. Werecommend all users of QIAGEN products to adhere to the NIH guidelines that have beendeveloped for recombinant DNA experiments, or to other applicable guidelines.

    Product Warranty and Satisfaction GuaranteeQIAGEN guarantees the performance of all products in the manner described in our productliterature. 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, QIAGENwill replace it free of charge or refund the purchase price. We reserve the right to change,alter, or modify any product to enhance its performance and design. If a QIAGEN productdoes not meet your expectations, simply call your local Technical Service Department ordistributor. We will credit your account or exchange the product as you wish.

    A copy of QIAGEN terms and conditions can be obtained on request, and is also providedon the back of our invoices. If you have questions about product specifications or performance, please call QIAGEN Technical Services or your local distributor (see backcover).

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  • QIAamp DNA Blood Midi/Maxi Handbook 04/20106

    Technical AssistanceAt QIAGEN we pride ourselves on the quality and availability of our technical support.Our Technical Service Departments are staffed by experienced scientists with extensivepractical and theoretical expertise in molecular biology and the use of QIAGEN products.If you have any questions or experience any difficulties regarding QIAGEN DNA BloodMidi or Maxi Kits or QIAGEN products in general, please do not hesitate to contact us.

    QIAGEN customers are a major source of information regarding advanced or specializeduses of our products. This information is helpful to other scientists as well as to theresearchers at QIAGEN. We therefore encourage you to contact us if you have any suggestions about product performance or new applications and techniques.

    For technical assistance and more information please call one of the QIAGEN TechnicalService Departments or local distributors (see back cover).

    Quality ControlAs part of the stringent QIAGEN quality assurance program, the performance of QIAampDNA Blood Midi and Maxi Kits is monitored routinely on a lot-to-lot basis. All componentsare tested separately to ensure highest performance and reliability.

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  • QIAamp DNA Blood Midi/Maxi Handbook 04/2010 7

    Safety InformationWhen working with chemicals, always wear a suitable lab coat, disposable gloves, andprotective goggles. For more information, please consult the appropriate material safetydata sheets (MSDSs). These are available online in convenient and compact PDF formatat www.qiagen.com/ts/msds.asp where you can find, view, and print the MSDS foreach QIAGEN kit and kit components.

    CAUTION: DO NOT add bleach or acidic solutions directly to the sample-preparation waste.

    The sample-preparation waste contains guanidine hydrochloride from Buffers AL andAW1, which can form highly reactive compounds when combined with bleach.

    If liquid containing these buffers is spilt, clean with suitable laboratory detergent andwater. If the spilt liquid contains potentially infectious agents, clean the affected area firstwith laboratory detergent and water, and then with 1% (v/v) sodium hypochlorite.

    The following risk and safety phrases apply to components of the QIAamp DNA BloodMidi and Maxi Kits:

    Buffers AW1 and AL

    Contain guanidine hydrochloride: harmful, irritant. Risk and safety phrases:*R22-36/38, S13-26-36-46

    QIAGEN Protease

    Contains subtilisin: sensitizer, irritant. Risk and safety phrases:* R37/38-41-42, S22-24-26-36/37/39-46

    24-hour emergency information

    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 andskin; R41: Risk of serious damage to eyes; R42: May cause sensitization by inhalation; S13: Keep away fromfood, drink and animal feedingstuff; S22: Do not breathe dust; S24: Avoid contact with skin; S26: In case ofcontact 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 this container or label.

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  • QIAamp DNA Blood Midi/Maxi Handbook 04/20108

    IntroductionQIAamp DNA Blood Midi and Maxi Kits provide the fastest and easiest way to purify totalDNA for reliable PCR and Southern blotting. Total DNA (e.g., genomic, viral, mitochondrial)can be purified from up to 0.32 ml and 310 ml, respectively, of whole blood, plasma,serum, buffy coat, bone marrow, other body fluids, and lymphocytes.

    The simple QIAamp DNA Blood Midi and Maxi procedures yield pure DNA ready fordirect restriction digestion or amplification. With a suitable centrifuge r

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