\>ll :. Neasy B!c?od & Tissue KH DNeasy 96 BSood & Tissue Kit For purincaiion ol tota! DNA from animal biood animai isssue 9. vl t cultured cells ixe< » bacteria QIAGEN DOH-DL 15/16-02RT1I Document No. 1 RTI Release Trademarks: QiAGEN* DNeosy*, KNeasy* (QIAGEN Group); DU* (Beckman instrumenis. Inc.); hvacP (Matrix Technologies Corp.); Triton® (Rohm tKfd Haas Company). The PCR process is cowCTed by the foreign counterparts oi U.S. PcênfeNos, 4,683,202 cmd 4,683,195 owned by F. Hoffmann- La Roche Ltd. QtAzot lysis Recent is a subject of US Paîl No. 5,346,994 and for^gn equivaierds. -'RNA/atef®" is a âclemoTk of AMBiON, Inc., Au^m, Texas and is covered by vunows U.S. and foreign patents. 8 2006 QIAGEN. a8 ligfits reserved. DOH-DL 15/16-02RT1I Document No. 2 RTI Release 5 Sfsraga & ?FOt'SiCt 6JS3 I-!."a[tcfeRS Product WaitiBiity and Sciiismeiiati ©stdffaniae Taciinicai AM;sScnce ,<**^ _ _B* iyh 8 9 Principle and procedure 9 Description of protocois 12 EquipsionS cetd Raagsats I® E® Sug^risd try Ussr 13 impoE'tan'? êSas S5 Sample colection and storage 15 Starting amounts of samples 15 Maximum amouro of starting material 15 Very smal sampie sizes 15 QuanNftcation of starting material 17 Preparation of Buffer AW 1 and Buffer AW2 18 Buffer AL 18 Proteinase K 19 Copurification of RNA 19 Centrifugation (DNeasy 96 procedures) 20 Elution of pure nucieic acids 21 Expected yields 22 Purification of high-molecular-weight DNA ftmSaaiio.i F'B'otnosfe .?. DSl;A yre'm "inmiaS »31^. 'S.'CBI AniiB'a! TTssues (Sriin-C?^'By°<'wFS.wi :?«'<»S'oci^j 28 Bbarf ay Cs^s isasy 96 ?n;(ceo!) 31 &gi5on Q& "EnSaE DMA {TOWI DNeasy Blood & Tissue Handbook 07/2006 3 DOH-DL 15/16-02RT1I Document No. 3 RTI Release iToioaois Preireo&nsn!" foi- Po.';aSa-!.:inb®ilei8(J 'fissua y.'i Preis'esatmen!' f®j' Fo.'ffiiaiin-^sioti Tissii® (r:j.'^" Qflam-Megafr.?® A4 PrehôEmsnS' for Snsm-PcsiSv® SccSeric .^ "s'i-oublesliooiiag ©uide 4;7' A^pen^'ix A: DateniiiaoSaoii c^ViAl, ?uriiy, and S.&igA; ®f 9iÂ >*^. Appsn&ci B: Cieenmg S-BSesSts 54, Qi-dsring InScfmoiion .55 Q!AG?:M Disi'ribssters and imsa?'2"i's W1 DNeasy Blood & Tissue Handbook 07/2006 DOH-DL 15/16-02RT1I Document No. 4 RTI Release rt,-T°" .-^ . -i . . r. 5i\^ inJ^* 'l'" ^(.j^. iL^;ir'U^j'\?i~''?iir .J"Ê< &11ssusKEB- Nurabsr sS p»®§is 50 250 DNeasy Mini Spin Columns (coloHess) in 2 ml Colection Tubes 50 250 Colection Tubes j2 ml) 100 500 Buffer ATL 10ml 50ml Buffer AL* 12ml 54ml BuferAWI (concenh-ate)*' 19ml 95 ml Buffer AVV2 (concentrate)" i 3 ml 66 mi Buffer AE 22m! 2 x 60 ml Protainase K ? 1.25 ml 6m! 1 Contains a chaoftopic salt. Not compaSbte with disinfed'ng mts asntalning bleach. See |xige 8 For safety informction. Buffer AW 1 and Buifer AW2 are 5uppSied as concentrates. ethanol (96-100%) according to the bottle labei before use to obtain c woHclng soluôn. Conhains sodium azide as a presercafive. DNeasy Blood & Tissue Handbook 07/2006 5 DOH-DL 15/16-02RT1I Document No. 5 RTI Release DNeas"- 96 3icod & Tisus Ki? w CtSiWag no. !1 592 N'cniber e& preps 4x96 12x% DNeasy 96 Plates 4 12 S-Blocks* 2 2 Colection Microtubes, 1.2 ml (racked) 4x96 12x96 Collection Microtube Caps 2xp2©%%5x(»2Qx8} Elution Microtubes RS (racked) and caps 4x96 12x96 AirPore Tape Sheets 25 3x25 Buffer AL* 86ml 247ml Buffer ATI 80ml 3 x 80 ml Buffer AV\/1 (concentratej" 98ml 3 x 98 ml Buffer AW2 (concentrate)'5 68 mi 3 x 68 ml Buffer AE 2x 110ml 500ml Proteinase K 2 x 7 m! 5 x 7 ml 96-Weil-Plate Register 4 12 Handbook 1 Reusable; see Appendix B (page 54} for leaning instructions. Contains a chootropic salt. Not compatible with disinfectanfs containing bleach. See page 8 for safety information. Buffer AW1 and Buffer AW2 are supplied as concenlralas. Add ethanol ;96-100%) according to the bottle label before use to obtain a working scûtkwi. Contains sodium azide as a preservative. .sitec^gi® DNeasy spin columns, DNeasy 96 plates, and all buffers should be stored dry, at room temperature (15-25°C) and are stable for 1 year under these conditions. DNeasy Blood & Tissue Kits contain a ready-to-use proteinase K solution, which is supplied in a specially formulated storage buffer. Proteinase K is stable for at least 1 year after delivery when shared at room temperature. For storage longer than one year or if ambient temperatures often exceed 25°C, we suggest storing proteinase K at 2-8°C. 6 DNeasy Blood & "nssue Handbook 07/2006 DOH-DL 15/16-02RT1I Document No. 6 RTI Release DNeasy Blood & Tissue Kits and DNeasy 96 Blood & Tissue Kits are intended for research use. No claim or representation is intended to provide information 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. ^FLT' l-C'LVX'^''^/r'^.;"T- ..K7>.-jlu'T'u>.l'r~:'t &5'l'J©?BC'li QIAGEN guarantees the performance of all products in the manner described in our product literature. Vne purchaser inust determine the suitability of the product for its particular use. Should any product fail to perform satisfactorily due to any reason other than misuse, QSAGEN will replace it free of charge or refund the purchase price. We reserve the right to change, alter, or modify ony product to enhance its performance and design. If a QIAGEN product does not meet your expectations, simply ca!l 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 caii QIAGEN Technical Services or your locai distributor (see back coveri. ^ .-.r1^ _.*.-. -.... H .^ -. _° -.^- Jl-i-":?y'^^tJLvilvjliiMJ'*Ari:" .'- L\; \i. -;frf'A\^''Sv»frL.J"*:^w.:l.:7^LK>-^:-'("i1;^'?i At QIAGEN we pride ourselves on the quaisfy 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 difficuities regarding DNeasy Blood & Tissue Kits or QIAGEN products in general, please do not hesitate to contact us. QIAGEN customers are a major source of informah'on regarding advanced or specialized uses of our products. This information is helphil to other scientists as well as to th® ressarchsrs at Q!AGEN. Wa therefcrs sncouragB you ?o contact us if you havs any suggestions abou^ product performance or new appiications and techniques. For technical assistance and more information please call one of the QIAGEN Technical Service Departments or beat distributors {see back cover). DNeasy Blood & Tissue Handbook 07/2006 DOH-DL 15/16-02RT1I Document No. 7 RTI Release @y'3sii;y ^.^.©1 tl' In accordance with QSAGEN's !SO<:ertified Quality Management System, each lot of DNeasy Blood & Tissue Kits and DNeasy 96 Biood & Tissue Kits is tested against predetermined specifications to ensure consistent product quality. .^ ^ . .: n 'f- - -- "c ^_':x1^,'?.~^*h'-^ ";~yV j->\7( '_,;r.~l^'.i :i^^(7-1Lr?:r-^,'^,i'tf->;-'jI^7i:l'^T)'i.)rj^j1 When working with chemicals, always wear a suitable lab coat, disposable gloves, and protective goggles. For more informaKon, please consult the appropriate material safety data sheets (MSDSsj. These are ayaitabie online in convenient and compact PDF format at www.qiaaen.com/ts/msds.asp where you can find, view, and print the MSDS for each QIAGEN kit and kit component. . eesdic soiuHsmsMsetSy ic fhe s

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