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Technical BulletinWizard®SVGel and PCRClean-Up SystemINSTRUCTIONS FOR USE OF PRODUCTS A9280, A9281 ANDA9282.PRINTED IN USA Revised 1/05AF9TB308 0105TB308Part# TB308Wizard®SV Gel and PCR Clean-Up SystemAll technical literature is available on the Internet at www.promega.com/tbs Please visit the web site to verify that you are using the most current version of thisTechnical Bulletin. Please contact Promega Technical Services if you have questions on useof this system. E-mail techserv@promega.com.I.II.III.IV.Description..........................................................................................................1Product Components.........................................................................................3General Considerations....................................................................................4Gel Slice and PCR Product Preparation........................................................4A.Preparing the Membrane Wash Solution.........................................................4B.Dissolving the Gel Slice.......................................................................................5C. Processing PCR Reactions...................................................................................6V.DNA Purification...............................................................................................6A.DNA Purification by Centrifugation.................................................................6B.DNA Purification by Vacuum............................................................................7VI.Troubleshooting.................................................................................................9VII.References.........................................................................................................11VIII.Appendix...........................................................................................................11A.Composition of Buffers and Solutions............................................................11B. Related Products.................................................................................................12I.DescriptionThe Wizard®SV Gel and PCR Clean-Up System is designed to extract andpurify DNA fragments of 100bp to 10kb from standard or low-melt agarose gelsin either Tris acetate (TAE) or Tris borate (TBE), or to purify PCR productsdirectly from a PCR amplification(a). Up to 95% recovery is achieved dependingupon the DNA fragment size (see Table 1). PCR products are commonlypurified to remove excess nucleotides and primers. This membrane-basedsystem, which can bind up to 40µg DNA, allows recovery of isolated DNAfragments or PCR products in as little as 20 minutes, depending on the numberof samples processed and the protocol used. The purified DNA can be used forautomated fluorescent DNA sequencing, cloning, labeling, restriction enzymedigestion or in vitro transcription/translation without further manipulation.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPrinted in USA.Revised 1/05Part# TB308Page 1The Wizard®SV Gel and PCR Clean-Up System is based on the ability of DNA tobind to silica membranes in the presence of chaotropic salts. After electrophoresisto separate the DNA fragments, the band(s) of interest is excised and dissolvedin the presence of guanidine isothiocyanate (Membrane Binding Solution).Alternatively, after amplification, an aliquot of PCR reaction is added to theMembrane Binding Solution and directly purified. The system allows a choice ofmethods for isolation of DNA from the dissolved agarose gel slice or PCRreaction. DNA can be isolated using microcentrifugation to force the dissolvedgel slice or PCR reaction through the membrane while simultaneously bindingthe DNA on the surface of the silica (Section V.A). After washing the isolatedDNA fragment or PCR product, the DNA is eluted in water. Another option ispulling the dissolved gel or PCR reaction through the SV Minicolumn andwashing the DNA fragment using vacuum pressure (Section V.B). The VacuumAdapters allow the use of a vacuum manifold (e.g., Vac-Man®LaboratoryVacuum Manifold, 20-sample capacity [Cat.# A7231], or Vac-Man®Jr. LaboratoryVacuum Manifold, 2-sample capacity [Cat.# A7660]). The Vacuum Adapters(Cat. # A1331) are only supplied with Cat.# A9280, Wizard®SV Gel and PCRClean-Up System, 10 preps, but may be purchased separately.The Wizard®SV Gel and PCR Clean-Up System can be used with linear DNAfragments, supercoiled plasmid DNA, or single-stranded linear or circular DNA.Expected yields with single-stranded DNA are lower than for double-strandedDNA.Table 1. Percent Recovery VersusDouble-Stranded DNA Fragment Size.PCR products (55–1,000bp), linearizedpGEM®-3Zf(+) plasmid (3,199bp), orLambda Hind III fragments (9,416bp and23,130bp) were purified in triplicate froma 1% agarose gel slice in 1X TAE bufferand quantified by ethidium bromidestaining.DNA Fragment SizePercent Recovery55bp70bp85bp100bp500bp1,000bp3,199bp9,416bp23,130bp26%39%55%84%%92%95%95%47%Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPart# TB308Page 2Printed in USA.Revised 1/05II.Product ComponentsProductSizeCat. #®WizardSV Gel and PCR Clean-Up System10 prepsA9280 For Laboratory Use. Each system contains sufficient reagents for 10 purifications.Includes:•••••••4ml3ml1.25ml101051Membrane Binding SolutionMembrane Wash Solution (concentrated)Nuclease-Free WaterWizard®SV MinicolumnsCollection Tubes (2ml)Vacuum AdaptersProtocolProductSizeCat. #®WizardSV Gel and PCR Clean-Up System50 prepsA9281 For Laboratory Use. Each system contains sufficient reagents for 50 purifications.Includes:••••••20ml15ml3.75ml50501Membrane Binding SolutionMembrane Wash Solution (concentrated)Nuclease-Free WaterWizard®SV MinicolumnsCollection Tubes (2ml)ProtocolProductSizeCat. #®WizardSV Gel and PCR Clean-Up System250 prepsA9282 For Laboratory Use. Each systemcontainssufficient reagents for 250purifications.Includes:••••••100ml75ml13ml2502501Membrane Binding SolutionMembrane Wash Solution (concentrated)Nuclease-Free WaterWizard®SV MinicolumnsCollection Tubes (2ml)ProtocolStorage Conditions:Store all components at room temperature (22–25°C). Norefrigeration is required. Keep Membrane Binding Solution protected fromlight. See expiration date on product label.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPrinted in USA.Revised 1/05Part# TB308Page 3III.General ConsiderationsAgarose, a linear polymer extracted from seaweed, is commonly used forelectrophoretic separation of nucleic acids. Standard agarose melts at 87–°C andsolidifies at 36–39°C. In low-melt agarose, hydroxyethyl groups have beenintroduced into the polysaccharide chain, resulting in an agarose that both meltsand solidifies at much lower temperatures (65°C and 24–28°C, respectively). Low-melt agarose is often used for applications that require recovery of intact DNAfragments from the gel after electrophoresis. The Wizard®SV Gel and PCR Clean-Up System can be used to recover DNA from either standard or low-melt agarosegels with no changes to the protocol or differences in recovery (Section V).Standard safety apparel should be worn, especially when handling ethidiumbromide-stained agarose gels. This includes gloves and a UV-blocking face shieldto protect the eyes and face from UV light. When excising the gel band, workquickly to minimize personal exposure to UV light and to minimize nicking of theDNA (1–4).The Wizard®SV Gel and PCR Clean-Up System is compatible with PCR productsgenerated using a variety of amplification enzymes, buffers or PCR-enhancingadditives. Mineral oil does not interfere with purification.IV.Gel Slice and PCR Product PreparationMaterials to Be Supplied by the User(Solution compositions are provided in SectionVIII.A.)••••••A.1.5ml microcentrifuge tubesethanol (95%)Vacuum Adapters (Cat.# A1331; only for vacuum purification)agarose gel (standard or low-melt; only for gel purification)1X TAE or TBE electrophoresis buffer (only for gel purification)50–65°C heating block (only for gel purification)Preparing the Membrane Wash SolutionAdd the indicated volume of 95% ethanol to the Membrane Wash Solution priorto beginning the procedure (see Table 2). Mark the bottle label to record that thisaddition was made. Tightly close the bottle cap after each use to preventevaporation.Table 2. Volume of 95% Ethanol to Add to Membrane WashSolution for Each System Size10 preps50 preps250 prepsPart Number ofVolume of 95%Membrane Wash SolutionEthanolSystem Size.A929AA929BA929C15ml75ml375mlPromega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPart# TB308Page 4Printed in USA.Revised 1/05B.Dissolving the Gel Slice1.Load and run the gel using an established protocol. DNA can be extractedfrom standard or low-melt agarose gels run with either TAE or TBE buffer.2.Weigh a 1.5ml microcentrifuge tube for each DNA fragment to be isolatedand record the weight.3.Visualize and photograph the DNA using a long-wavelength UV lamp andan intercalating dye such as ethidium bromide. To reduce nicking, irradiatethe gel for the absolute minimum time possible (1–4). Excise the DNAfragment of interest in a minimal volume of agarose using a clean scalpelor razor blade. Transfer the gel slice to the weighed microcentrifuge tubeand record the weight. Subtract the weight of the empty tube from the totalweight to obtain the weight of the gel slice (see Notes 1–3 below).Note: The gel slice may be stored at 4°C or at –20°C for up to one week in atightly closed tube under nuclease-free conditions before purification.4.Add Membrane Binding Solution at a ratio of 10µl of solution per 10mg ofagarose gel slice.5.Vortex the mixture (see Note 4) and incubate at 50–65°C for 10 minutes oruntil the gel slice is completely dissolved. Vortex the tube every few minutesto increase the rate of agarose gel melting. Centrifuge the tube briefly atroom temperature to ensure the contents are at the bottom of the tube. Oncethe agarose gel is melted, the gel will not resolidify at room temperature.6.To purify the DNA using a microcentrifuge, proceed to Section V.A. Topurify the DNA using a vacuum manifold, proceed to Section V.B.Notes:1.Recovery from 1% high-melting-point agarose is comparable to that from1–2% low-melting-point agarose. High-melting-point agarose concentrationsof up to 3% have been tested. Gel slices with higher agarose concentrations(2–3%) may require a longer time to melt completely than a 1% agarose gelslice and may show reduced yields.2.The maximum capacity of the column is 350mg of gel mass dissolved in350µl of Membrane Binding Solution per column pass. For gel slices >350mg,continue to pass additional sample through the SV Minicolumn until all ofthe sample has been processed. The maximal amount of agarose that can beprocessed through a single column is approximately 3.5g (10 × 350mg) total.3.The maximum binding capacity of the column is approximately 40µg percolumn, and as little as 10ng has been successfully purified.4.DNA fragments that are larger than 5kb should be mixed gently to preventshearing. Do not vortex if DNA fragment is larger than 5kb; mix byinversion.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPrinted in USA.Revised 1/05Part# TB308Page 5C.Processing PCR Reactions1.Amplify target of choice using standard amplification conditions.2.Add an equal volume of Membrane Binding Solution to the PCR reaction(see Notes 1–4 below).3.To purify the DNA using a microcentrifuge, proceed to Section V.A. To purify the DNA using a vacuum manifold, proceed to Section V.B.Notes:1.The maximal capacity of a single SV Minicolumn is approximately 1ml ofPCR reaction added to 1ml Membrane Binding Solution (2ml total). ForPCR volumes >350µl, continue to pass the sample through the column untilall of the sample has been processed.2.The maximum binding capacity is approximately 40µg per column, and aslittle as 10ng has been successfully purified.3.Mineral oil does not interfere with purification.4.For amplification reactions that do not produce a single product or whereamplification has been inefficient and there is highly visible primer dimer,gel purification of the band of interest is recommended. Alternatively, an80% ethanol wash solution can be substituted for the supplied MembraneWash Solution to reduce primer-dimer carryover.V.DNA PurificationPrepare the gel slice or PCR product as described in Section IV. Use either thecentrifugation procedure (Section V.A) or the vacuum procedure (Section V.B) torecover the DNA from the dissolved gel slice or PCR reaction. After theprocedure is completed, the DNA may be used in downstream applications.A.DNA Purification by Centrifugation1.Place one SV Minicolumn in a Collection Tube for each dissolved gel sliceor PCR reaction.2.Transfer the dissolved gel mixture or prepared PCR product to the SVMinicolumn assembly and incubate for 1 minute at room temperature.3.Centrifuge the SV Minicolumn assembly in a microcentrifuge at 16,000 × g(14,000rpm) for 1 minute. Remove the SV Minicolumn from the SpinColumn assembly and discard the liquid in the Collection Tube. Return theSV Minicolumn to the Collection Tube.!Note: Failure to spin at 16,000 × g(14,000rpm) can result in reduced yield.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPart# TB308Page 6Printed in USA.Revised 1/0.Wash the column by adding 700µl of Membrane Wash Solution, previouslydiluted with 95% ethanol (see Section IV.A), to the SV Minicolumn.Centrifuge the SV Minicolumn assembly for 1 minute at 16,000 × g(14,000rpm). Empty the Collection Tube as before and place the SVMinicolumn back in the Collection Tube. Repeat the wash with 500µl ofMembrane Wash Solution and centrifuge the SV Minicolumn assembly for5 minutes at 16,000 × g.5.Remove the SV Minicolumn assembly from the centrifuge, being careful notto wet the bottom of the column with the flowthrough. Empty the CollectionTube and recentrifuge the column assembly for 1 minute with the micro-centrifuge lid open (or off) to allow evaporation of any residual ethanol.6.Carefully transfer the SV Minicolumn to a clean 1.5ml microcentrifuge tube.Apply 50µl of Nuclease-Free Water directly to the center of the columnwithout touching the membrane with the pipette tip. Incubate at roomtemperature for 1 minute. Centrifuge for 1 minute at 16,000 × g(14,000rpm).7.Discard the SV Minicolumn and store the microcentrifuge tube containingthe eluted DNA at 4°C or –20°C.Note:The volume of the eluted DNA will be approximately 42–47µl. If theDNA needs to be further concentrated, perform an ethanol precipitation.Alternatively, the DNA may be eluted in as little as 15µl of Nuclease-FreeWater without significant reduction in yield. If using an elution volume of15µl, verify that the membrane is completely covered with Nuclease-FreeWater before centrifugation. Elution volumes less than 15µl are notrecommended (see Table 3).B.DNA Purification by Vacuum1.Attach one Vacuum Adapter with a Luer-Lok®fitting to one port of themanifold (e.g., Vac-Man®or Vac-Man®Jr. Laboratory Vacuum Manifold)for each dissolved gel slice or PCR reaction. Insert SV Minicolumn into theVacuum Adapter until it fits snugly in place.2.Transfer the dissolved gel mixture or PCR reaction to the SV Minicolumnand incubate for 1 minute at room temperature. Apply a vacuum to pullthe liquid completely through the SV Minicolumn.Note:The minimum vacuum pressure is 15inches of mercury. See the table to the rightfor comparison of inches of Hg to otherpressure measurements.1 Inch Hg3.386kPa25.4Torr0.0334atm0.491psi2.cm Hg33.86mbar15 Inches Hg50.8kPa381Torr0.501atm7.37psi38.1cm Hg508mbarPromega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPrinted in USA.Revised 1/05Part# TB308Page 73.Wash the column by adding 700µl Membrane Wash Solution previouslydiluted with 95% ethanol (see Section IV.A) to the SV Minicolumn. Make sureany droplets remaining on the sides of the SV Minicolumn from the last stepare washed away. Apply a vacuum to pull the liquid through the SVMinicolumn. Repeat this wash a second time with 500µl of Membrane WashSolution.4.Turn off the vacuum source and open an unused port to vent the manifold.Remove the SV Minicolumn from the vacuum manifold and transfer to aCollection Tube. Centrifuge the SV Minicolumn assembly for 5 minutes at16,000 × g(14,000rpm) to remove any remaining Membrane Wash Solution.5.Empty the Collection Tube and recentrifuge the column assembly for 1 minutewith the microcentrifuge lid open (or off) to allow evaporation of any residualethanol.6.Carefully transfer the SV Minicolumn to a clean 1.5ml microcentrifuge tube,being careful not to wet the bottom of the SV Minicolumn with theflowthrough. Apply 50µl of Nuclease-Free Water directly to the center of thecolumn without contacting the membrane. Incubate at room temperature for 1 minute. Centrifuge for 1 minute at 16,000 × g(14,000rpm).7.Discard the SV Minicolumn and store the microcentrifuge tube containing theeluted DNA at 4°C or –20°C.Note:The volume of the eluted DNA will be approximately 42–47µl. If the DNAneeds to be further concentrated, perform an ethanol precipitation. Alternatively,the DNA may be eluted in as little as 15µl of Nuclease-Free Water without asignificant reduction in yield. If using an elution volume of 15µl, verify that themembrane is completely covered with Nuclease-Free Water before centrifugation.Elution volumes less than 15µl are not recommended (see Table 3).Table 3. Percent Recovery VersusElution Volume. A 700bp PCR productwas directly purified in triplicate andquantified by ethidium bromide staining.Elution Volume10µl15µl25µl50µl75µl100µlPercent RecoveryCompared to 50µl35%98%98%100%100%100%Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPart# TB308Page 8Printed in USA.Revised 1/05VI.TroubleshootingFor questions not addressed here, please contact your local Promega Branch Office or Distributor.Contact information available at: www.promega.com. E-mail: techserv@promega.comSymptomsLow DNA yieldCauses and CommentsVerify that an equal volume of Membrane Binding Solution was added to the gel slice or PCR (10µl per 10mg gel slice or 10µl PCR).Make certain that the gel slice is completely melted before proceeding with the purification. Incubation at 50–65°C is necessary to completely melt the gel slice.If the amount of DNApurified is too small to quantitate by spectrophotometry, quantitate by agarose gel electrophoresis followed by ethidium bromide or PicoGreen®staining.Be sure to centrifuge at 16,000 × g(14,000rpm).Verify that ethanol was added to the MembraneWash Solution (see Section IV.A) and repeat thepurification.Poor results with automated fluorescent sequencingToo little DNAmay have been used. Increase theamount of DNA used in sequencing reactionsor concentrate the DNA by ethanol precipitation.Up to 7µl of the eluted DNA can be used per fluorescent sequencing reaction.Too much DNA can interfere with fluorescent sequencing. Use less eluted DNA or dilute DNA prior to sequencing.If TE was used for elution, ethanol precipitate the DNA or repurify the DNA fragments and elute with Nuclease-Free Water.Excessive thymidine-dimer formation may haveoccurred during UV exposure. See references 1–4 for a method to minimize thymidine-dimer formation of AT-rich templates.Poor restriction digestionIncrease the amount of restriction enzyme and/or the length of incubation time. Digest at the appropriate temperature and in the optimal buffer for the restriction enzyme used.Ethanol or salt carryover into the eluted DNAmay have occurred. Ethanol precipitate the DNA or keep the DNA volume to 10% or less ofthe final reaction volume.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPrinted in USA.Revised 1/05Part# TB308Page 9VI.Troubleshooting (continued)SymptomsDNA yields on gel look lowcompared to spectrophotometricreadingsCauses and CommentsTrace contaminants in eluted DNA can artificiallyinflate spectrophotometer readings. Use agarosegel electrophoresis followed by ethidium bromideor PicoGreen®staining to determine DNA yields.Ethanol precipitate the DNA.Low A260/A230ratiosTypically due to guanidine isothiocyanate contamination. Low ratios do not necessarily indicate that the DNA will function poorly in downstream applications. Ethanol precipitate the DNA if low A260/A230ratio is a concern.Increase the length of the 50–65°C incubation to ensure the gel slice is completely melted.Verify that an equal ratio of Membrane Binding Solution to gel slice mass is used (10µl per 10mg). A vacuum pressure of >15 inches of mercury is required to use the SV Minicolumn in the vacuum protocol. If the vacuum is insufficient, use the spin protocol.Purified DNA floats out of thewell when loaded on a gelEthanol carryover. Be certain that the MembraneWash Solution is not carried over from the wash steps. If the column has been wet, empty the Collection Tube and recentrifuge the column assembly for 1 minute. Centrifuge the SV Minicolumn for 5 minutes to remove residual Membrane Wash Solution. After washing, centrifuge the column assembly with the microcentrifuge lid open or off (Section V.A., Step 5; Section V.B., Step 5) to allow evaporation of any residual ethanol.Add 3X loading dye to the DNA sample before loading onto the gel.Purified DNA bands are not sharpDNAmay be sheared. Mix the agarose gel slice gently with the Membrane Binding Solution.Nuclease contamination may be an issue. Autoclave the gel running buffer before use.Store the gel slice at 4°C or –20°C for no more than 1 week under nuclease-free conditions. Low cloning efficiencyMay be due to guanidine isothiocyanate contamination. Ethanol precipitate the DNA, washing the pellet with 70% ethanol to reduce contamination.Clogged spin basketPromega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPart# TB308Page 10Printed in USA.Revised 1/05VII.References1.Zimmermann, M., Veeck, J. and Wolf, K. (1998) Minimizing the exposure to UV lightwhen extracting DNA from agarose gels. BioTechniques25, 586.2.Hengen, P. (1997) Methods and reagents. Protecting vector DNA from UV light.Trends Biochem. Sci.22, 182–3.3.Grundemann, D. and Schomig, E. (1996) Protection of DNA during preparativeagarose gel electrophoresis against damage induced by ultraviolet light. BioTechniques21, 8–903.4.Cariello, N.F. et al. (1988) DNA damage produced by ethidium bromide staining andexposure to ultraviolet light. Nucl. Acids Res.16, 4157.VIII.AppendixA.Composition of Buffers and SolutionsMembrane Wash Solution (after ethanol addition)10mMpotassium acetate (pH 5.0)80%ethanol16.7µMEDTA (pH 8.0)To prepare this solution, add 95%ethanol to the supplied MembraneWash Solution (concentrated) asdescribed in Table 2 in Section IV.A.Membrane Binding Solution4.5Mguanidine isothiocyanate0.5Mpotassium acetate (pH 5.0)1X TE buffer10mMTris-HCl (pH 7.5)1mMEDTA (pH 8.0)1X TBE buffermMTris basemMboric acid2mMEDTA (pH 8.0)1X TAE buffer40mMTris base5mMsodium acetate1mMEDTA (pH 8.0)Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPrinted in USA.Revised 1/05Part# TB308Page 11B.Related ProductsProductVacuum AdaptersWizard®SV 96 PCR Clean-Up System*Size Cat. #20 eachA13311 × 96 prepsA93404 × 96 prepsA93418 × 96 prepsA9342100 × 96 prepsA9345PCR Master Mix*100 reactions M7502 1,000 reactionsM7505®GoTaqDNA Polymerase*100 unitsM3001500 unitsM30052,500 unitsM3008Access RT-PCR System*20 reactionsA1260100 reactionsA1250500 reactionsA1280AccessQuick™ RT-PCR System*20 reactionsA1701100 reactionsA1702500 reactionsA1703Agarose, LE, Analytical Grade*100gV3121500gV3125Ethidium Bromide Solution, Molecular Grade*10mlH5041TAE Buffer,10X1,000mlV4271TBE Buffer, 10X1,000mlV4251*For Laboratory Use.(a)The PCR process is covered by patents issued and applicable in certain countries*.Promega does not encourage orsupport the unauthorized or unlicensed use of the PCR process.*In the U.S., effective March 29, 2005, U.S.Pat.Nos.4,683,195, 4,965,188 and 4,683,202 will expire.In Europe, effectiveMarch 28, 2006, European Pat.Nos.201,184 and 200,362 will expire.© 2002–2005 Promega Corporation.All Rights Reserved.GoTaq, Vac-Man and Wizard are registered trademarks of Promega Corporation.AccessQuick is a trademark of PromegaCorporation.Luer-Lok is a registered trademark of Becton, Dickinson and Company.PicoGreen is a registered trademark of MolecularProbes, Inc.Products may be covered by pending or issued patents or may have certain limitations.Please visit our Web site for moreinformation.All prices and specifications are subject to change without prior notice.Product claims are subject to change.Please contact Promega Technical Services or access the Promega online catalog forthe most up-to-date information on Promega products.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·www.promega.comPart# TB308Page 12Printed in USA.Revised 1/05