Bio-Spin-6-column.pdf

Bio-Spin-6-column.pdf

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nucleicacidpurificationBio-Spin6and30ColumnsAutoclavabilityBio-Spincolumns,Bio-GelPgel,andcollectiontubesarecompletelyautoclavableat121Cfor30minatpH6.08.0.ChemicalStabilitypH210,commonaqueousbuffers,formamide,diluteorganicacids,alcohol,20%（v/v）otherchaotropicagents,detergents.StorageStoreat4C.Donotfreeze.InstructionsforUse1Invertthecolumnsharplyseveraltimestoresuspendthesettledgelandremoveanybubbles.2Snapoffthetipandplacecolumnina2.0mlmicrocentrifugetube（included）.Removecap.Allowtheexcesspackingbuffertodrainbygravitytotopofgelbed.（Ifcolumndoesnotbegintoflow,pushcapbackintocolumnandremove）.Discardthedrainedbuffer,thenplacethecolumnbackinto2mltube.However,ifplacingcolumninto12x75mmtesttube,centrifugeimmediately.3Centrifugefor2mininaswingingbucketcentrifugeat1,000xg（seeCentrifugationNotessection）toremovethepackingbuffer.Discardthebuffer.4Placethecolumninaclean2.0mlmicrocentrifugetubeor12x75mmtesttube.Carefullyapplythesample（20100l）directlytothecenterofthecolumn.Applicationofmoreorlessthantherecommendedsamplevolumemaydecreasecolumnperformance.5Afterloadingsample,centrifugethecolumnfor4minat1,000xg.6Followingcentrifugation,thepurifiedsampleisnowinTrisorSSCbuffer.Moleculessmallerthanthecolumnsexclusionlimitwillberetained.7Properlydisposeoftheusedcolumn.BufferExchangeThegelintheBio-SpincolumnsissuspendedineitherSSCbuffer,pH7.0,orTrisbuffer,pH7.4.Thegelmatrixiscompatiblewithmostaqueousbuffers.Bufferexchangecanbeachievedusingthefollowingprocedure.IntroductionBio-Spinchromatographycolumnsarereadytouseforrapidandefficientcleanupandpurificationofnucleicacidsandproteinsusingaswingingbucketcentrifuge.Bio-Spin6and30ColumnsRemovedyeterminatorsRemoveunincorporatednucleotidesDesaltingandbufferexchangeThecolumnsarepackedwithspecialgradesofBio-GelPpolyacrylamideP-6orP-30gelmatricesmanufacturedspecificallyforBio-Radspincolumns.Thisuniquegelproducesveryefficient,noninteractivesizeseparations.WerecommendBio-Spin6Triscolumnsforbufferexchangeanddesaltingapplications,whileBio-Spin30Triscolumnsareoptimalforremovalofunincorporatednucleotides.Bio-Spincolumnsaresuitableforusewith2.0mlmicrocentrifugetubesor12x75testtubesandarecompletelyautoclavable.TechnicalInformationGelMatrixBio-GelP-6orP-30polyacrylamidegelsuspendedin1.0mlofbuffer.BuffersSSCbuffer（150mMsodiumchloride,17.5mMsodiumcitrate,pH7.0）with0.02%sodiumazide.Trisbuffer（10mMTris-HCl,pH7.4）with0.02%sodiumazide.SampleApplicationVolumesNucleicacids,proteins,andpeptides,20100l.ExclusionLimitsBio-GelP-6gel:

5basepairs（nucleicacids）ormolecularweight6,000（proteins,peptides）.Bio-GelP-30gel:

20basepairs（nucleicacids）ormolecularweight40,000（proteins,peptides）.ExpectedRetentionandRecoveryUpto99%retentionofunincorporatednucleotides.Upto95%recoveryofappliedDNA.CentrifugeTypeSwingingbucketcentrifugewithacentrifugalforceof1,000xg.LifeScienceGroupWebSitewww.bio-U.S.（800）4BIORADAustralia0299142800Austria（01）-8778901Belgium09-3855511Canada（905）712-2771China86-10-62051850/51Denmark4539179947Finland358（0）98042200France0143904690Germany08931884-0HongKong852-2789-3300India（91-11）461-0103Israel039514127Italy39-02-216091Japan03-5811-6270Korea82-2-3473-4460LatinAmerica305-894-5950Mexico514-2210TheNetherlands0318-540666NewZealand64-9-4152280Norway22-74-18-70Russia70959799800Singapore65-2729877Spain34-91-661-7085Sweden46（0）8-55512700Switzerland01-8095555UnitedKingdom0800-181134LIT-507RevGBio-RadLaboratories1Followsteps1,2,and3intheInstructionsforUsesection.2Applythenewbufferin500laliquots.Aftereachapplicationofnewbuffer,letthebufferdrainoutbygravity,orcentrifugethecolumnfor1mintoremovethebuffer.Discardbufferfromcollectiontube.Repeatasrequired.Threewashesresultin99%ofthebufferbeingexchanged.Fourwashesresultin99.9%ofbufferexchanged.3Samplecannowbeappliedtothecolumnasdirectedinsteps4through7intheInstructionsforUsesection.CentrifugationNotesBio-Spincolumnsfit2.0mlmicrocentrifugetubesor12x75mmtesttubesforsamplecollectionduringcentrifugation.Usethe2.0mlmicrotubesprovidedwiththecolumnsfortheinitialcolumnequilibrationstep.Swingingbucketcentrifugescapableofgeneratingaminimumforceof1,000xgaresuitableforBio-Spincolumnuse.Thegravitationalforcecreatedataparticularrevolutionspeedisafunctionoftheradiusofthemicrocentrifugerotor.Consulttheswingingbucketcentrifugeinstructionmanualforconversioninformationfromrevolutionsperminute（RPM）tocentrifugalorg-force.Alternatively,tocalculatethespeedinRPMrequiredtoreachagravitationalforceof1,000xg,usethefollowingequation:

RCF（xg）=（1.12x10-5）x（RPM）x2xrwhereRCFistherelativecentrifugalforce,ristheradiusincentimetersmeasuredfromthecenteroftherotortothemiddleoftheBio-Spincolumn,andRPMisthespeedoftherotor.SterilizationIfasterileBio-Spincolumnisrequired,autoclavethecolumnat121Cfor2030min.Ifexchangingbuffers,thebufferpHinthecolumnshouldbeintherangeof6.0to8.0priortoautoclaving.OrderingInformationCatalog#ProductDescriptionBio-SpinColumnswithBio-GelP-6inTrisBuffer732-6227Bio-Spin6TrisColumns,25732-6228Bio-Spin6TrisColumns,100732-6229Bio-Spin6TrisColumns,1,000Bio-SpinColumnswithBio-GelP-30inTrisBuffer732-6231Bio-Spin30TrisColumns,25732-6232Bio-Spin30TrisColumns,100732-6233Bio-Spin30TrisColumns,1,000Bio-SpinColumnswithBio-GelP-6inSSCBuffer732-6002Bio-Spin6SSCColumns,25Bio-SpinColumnswithBio-GelP-30inSSCBuffer732-6006Bio-Spin30SSCColumns,25MicroBio-SpinColumnswithBio-GelP-6inTrisBuffer732-6221MicroBio-Spin6TrisColumns,25732-6222MicroBio-Spin6TrisColumns,100732-6225MicroBio-Spin6TrisColumns,1,000MicroBio-SpinColumnswithBio-GelP-30inTrisBuffer732-6223MicroBio-Spin30TrisColumns,25732-6224MicroBio-Spin30TrisColumns,100732-6226MicroBio-Spin30TrisColumns,1,000MicroBio-SpinColumnswithBio-GelP-6inSSCBuffer732-6200MicroBio-Spin6SSCColumns,25732-6201MicroBio-Spin6SSCColumns,100MicroBio-SpinColumnswithBio-GelP-30inSSCBuffer732-6202MicroBio-Spin30SSCColumns,25732-6203MicroBio-Spin30SSCColumns,100RNase-FreeMicroBio-SpinColumnswithBio-GelP-30inTrisBuffer732-6250MicroBio-Spin30TrisColumns,RNasefree,25732-6251MicroBio-Spin30TrisColumns,RNasefree,100EmptyColumns732-6008EmptyBio-SpinColumns,100732-6204EmptyMicroBio-SpinColumns,100PCRReactionMixturePurification732-6300PCRKleenSpinColumns,25732-6301PCRKleenSpinColumns,100DNARecoveryfromAgaroseGels732-6165Freeze-N-SqueezeSpinColumns,25732-6166Freeze-N-SqueezeSpinColumns,100732-6160QuantumPrepGelSliceKitSEQueakyKleenTerminatorRemovalKit732-6260SEQueakyKleen,2x96732-6261SEQueakyKleen,10x96