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NEB/NEBNext® Library Quant Kit for Illumina®/E7630S/500 reactions
  • NEB/NEBNext® Library Quant Kit for Illumina®/E7630S/500 reactions

NEB/NEBNext® Library Quant Kit for Illumina®/E7630S/500 reactions

价格: ¥5040.00 市场价: 8400.00
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货号: E7630S-500reactions
品牌: NEB
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    • Description:











      TypicalresultsfromtheNEBNextLibraryQuantKitonaBio-RadCFX96Touch(A)andanAppliedBiosystems7500Fastreal-timeqPCRinstrument(B).Amplificationcurvesareshownontheleftandresultingstandardcurvesontheright.Alldefaultsettingswereusedonbothplatforms.ABI7500Fastassayincluded1XROX(LowConcentration)andtheROXnormalization.
      Accuratequantitationofnext-generationsequencinglibrariesisessentialformaximizingdataoutputandqualityfromeachsequencingrun.ForIlluminasequencingspecifically,accuratequantitationoflibrariesiscriticaltoachieveoptimalclusterdensities,arequirementforoptimalsequenceperformance.qPCRisconsideredtobethemostaccurateandeffectivemethodoflibraryquantitation,providingconsiderablyhigherconsistencyandreproducibilitythanelectrophoresisorspectrophotometry,whichmeasuretotalnucleicacidconcentration.Amplification-basedmethodsquantitateonlythosemoleculesthatcontainbothadaptorsequences,therebyprovidingamoreaccurateestimateoftheconcentrationofthelibrarymoleculesthatcanbesequenced.

      TheNEBNextLibraryQuantKitdeliverssignificantimprovementstoqPCR-basedlibraryquantitationfornextgensequencing.TheNEBNextLibraryQuantKitforIlluminacontainscomponentsthatareoptimizedforqPCR-basedquantitationoflibrariespreparedforIlluminanext-generationsequencingplatforms.TheNEBNextLibraryQuantKitcontainsprimerswhichtargettheP5andP7Illuminaadaptorsequencesandasetofhigh-quality,pre-dilutedDNAstandardstoenablereliablequantitationofdilutedDNAlibrariesbetween150–1000bp. 

      Features
      • ProvidesmoreaccurateandreproducIBLequantvaluesthanalternativemethodsandkits
      • CompatiblewithlibrarieswithabroadrangeofinsertsizesandGCcontent,madebyavarietyofmethods
      • Requiresonly4standards,allowingmorelibrariestobequantitatedperkit 
      • Suppliedwithaconvenient,LibraryDilutionBuffer
      • TheNEBNextLibraryQuantMasterMixrequiresonlytheadditionofprimers
      • Utilizesasingleextensiontimeforalllibraries,regardlessofinsertsize
      • LibraryquantvaluescanbeeasilycalculatedusingNEB’sonlinetool,atNEBioCalculator.neb.com
      • ROXisincludedinthekit,forusewithqPCRinstrumentsthatrequireareferencedyefornormalization
      NEBNextLibraryQuantKitworkflow NEBNext Library Quant Kit workflow
      qPCRprovidesmoreconsistentlibraryquantitationresultsthanBioanalyzer®analysis
      qPCR provides more consistent library quantitation results than Bioanalyzer® analysis

      Concentrationsof4librariesweredeterminedbytheNEBNextLibraryQuantKit(orange)andcomparedtovaluesmeasuredusingtheAglientBioanalyzer(blue).ComparedtoNEBNext’sqPCR-basedmethod,theBioanalyzerconcentrationsdisplayedagreaterlevelofvariation.

      GreaterreproducibilityoflibraryquantitationwiththeNEBNextLibraryQuantKit 
      Greater reproducibility of library quantitation with the NEBNext Library Quant Kit
      Three340–400bplibrarieswerequantitatedby4differentusers2–4timesusingeithertheNEBNextorKapaTMLibraryQuantificationKit(Universal).AnotableimprovementinquantitationconsistencywasobservedforconcentrationsdeterminedbytheNEBNextKit(orange)versusthosefromtheKapakit(gray). 
      Greaterlot-to-lotconsistencyofstandardswiththeNEBNextLibraryQuantKit 
      Greater lot-to-lot consistency of standards with the NEBNext Library Quant Kit
      AccurateqPCRquantitationrequirestheuseofhigh-qualityDNAstandardswithknownconcentrations.TheNEBNextLibraryQuantKitcontains4standardsproducedwithahighlevelofbothquantitationaccuracyandconsistency.Thisfigureshowsdatafrom>70totalrunsfrom4lotsofbothNEBNext(orange)andKapa(gray)standards,withallCqvaluesplotted.Boxandwhiskersindicatemeanandquartiles.TheNEBNextLibraryStandardsdisplayedmuchlowervariationinCq,resultinginmoreconsistentquantitationperformance. 
      TheNEBNextLibraryQuantKitvaluesenableoptimalclusterdensities
      The NEBNext Library Quant Kit values enable optimal cluster densities
      SevendifferentlibrarieswerequantitatedusingeithertheNEBNextLibraryQuantKit(orange)ortheKapaLibraryQuantificationKit(Universal)(gray).Undilutedlibraryconcentrationsrangedfrom2–200nM.Librariesweredilutedto8pMandloadedontoaMiSeq®instrument(v2chemistry;MCSv2.4.1.3).LibrariesquantitatedwiththeNEBNextkitresultedinarawclusterdensityaverageof1160k/mm2,directlyintheoptimalrangeof900–1300k/mm2.Incontrast,librariesloadedbasedontheKapaquantitationaveragedonly660k/mm2. 
      WiththeNEBNextLibraryQuantKit,optimalclusterdensityisachievedfromquantitatedlibrarieswithabroadrangeoflibrarysizeandGCcontent
      With the NEBNext Library Quant Kit, optimal cluster density is achieved from quantitated libraries with a broad range of library size and GC content
      Librariesof310–963bpfromtheindicatedsourceswerequantitatedusingtheNEBNextLibraryQuantKit,thendilutedto8pMandloadedontoaMiSeq(v2chemistry;MCSv2.4.1.3).Libraryconcentrationsrangedfrom7–120nM,andresultingrawclusterdensityforalllibrarieswas965–1300k/mm2(ave.=1199).OptimalclusterdensitywasachievedusingconcentrationsdeterminedbytheNEBNextLibraryQuantKitforalllibrarysizes. 
      AccurateLibraryQuantitationisachievedwithabroadrangeoflibrarysizeandGCcontent
      Accurate Library Quantitation is achieved with a broad range of library size and GC content
      AselectionoflibrariessuccessfullyquantitatedwiththeNEBNextLibraryQuantKit.Librariesareplottedbysize,rangingfromsmallerlibraries(sRNA,FFPE)at150–230bptolargestlibrariesat980bp.Variousinputsourceswereused,rangingfrom20-70%GCcontent,asindicatedbytextcolor.LibrarieswerepreparedusingNEBNext,IlluminaTruSeq®NanoandKapaHyperlibraryprepkits(datanotshown).NodependenceonsizeorGCcontentwasobservedinlibraryquantitationwhenusingNEBNext.

      LotControl
      ThelotsprovidedintheNEBNextLibraryQuantKitforIlluminaaremanagedseparatelyandarequalifiedbyfunctionaltesting.Individualreagentsundergostandardenzymeactivityandqualitycontrolassays,andalsomeetstringentcriteriaasdescribedoneachindividualcomponentpageinthemanual.

      FunctionallyValidated
      EachsetofreagentsisfunctionallyvalidatedtogetherthroughqPCR-basedlibraryquantitationassays.Reagentspassthefunctionaltestbyadheringtostringentcriteriasetforassayefficiencyandquantitationcycles(Cq)foreachDNAstandardandnotemplatecontrolreaction.

      Forlargervolumerequirements,customizedandbulkpackagingisavailablebypurchasingthroughtheOEM/BulksdepartmentatNEB.Pleasecontactnebsolutions@neb.comforfurtherinformation.

      ReagentsSupplied

      Thefollowingreagentsaresuppliedwiththisproduct:

      Storeat(°C)Concentration
      NEBNextLibraryQuantMasterMix-20
      NEBNextLibraryQuantPrimerMix-20
      NEBNextLibraryQuantDNAStandard1-20
      NEBNextLibraryQuantDNAStandard2-20
      NEBNextLibraryQuantDNAStandard3-20
      NEBNextLibraryQuantDNAStandard4-20
      ROX(Low)-20
      ROX(High)-20
      NEBNext®LibraryDilutionBuffer-2010X

      Notes:

      StorageofKitComponents:AlloftheindividualcomponentsoftheNEBNextLibraryQuantKitforIlluminaarestableat–20°Cfortwoyears.Whennotinuse,kitcomponentsshouldbestoredat–20°C.Allcomponentsofthekit,includingthecombinedNEBNextLibraryQuantMasterMixandPrimerMix,arestableforatleast30freeze/thaws.OncethePrimerMixhasbeenaddedtotheNEBNextLibraryQuantMasterMix(seeProtocolonpage6ofmanual),theresultingmixcanbestoredat–20°Cforsevenmonths.Forshort-termstorage,thismixmaybestoredat4°Cforuptosixweeks.Reactionscanbepreparedatroomtemperature,butunusedportionsofthekitreagentsshouldbekeptoniceafterthawingthemforuse.ForReagentsSupplied:Ifyouneeddilutionbufferinadditiontowhatisprovidedinthekit,itcanbepurchasedseparately.
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