MCLAB/MCNext™ SYBR® Fast qPCR Library Quantification Kit/IQPQRR/1 Ea

Description:
AccuratequantificationofNGSDNAlibrariesiscriticaltoensureefficientdatagenerationandhighqualityreads.QuantitativePolymeraseChainReaction(qPCR)isahighlysensitiveandaccurateapproachforquantifyingaNGSlibraryandusesaminimalamountofmaterialcomparedtootherquantificationmethods. TheqPCRquantificationtrackslibraryconcentrationsasafunction of PCR cycle numberin ordertoderive aquantitative estimateofthe initialtemplate concentrationbasedontheknownDNAconcentrationascontrolorstandard.Thus,thecontroltemplateshouldbeassimilaraspossIBLetothelibrariesforquantification,intermsoftemplatesize,GCcontentandlibrarytype.Itisalsoveryimportantthataconstantquantityofthecontroltemplateisavailable,asforeachroundoflibraryquantificationdeterminestheclustergenerationefficiency,inturntoadjusttheloADIngamountofconstructedlibrariesforthesequencing,evenslightvariationofthestandardleadstofluctuationofclusterdensity,eithercompromisethedataqualityorwastethesequencingcapacity. 

TheMCNext™SYBR^®FastqPCRLibraryQuantificationKitprovidesafastandreliablesolutiontodeterminethelibraryconcentrationwithadirectestimateclusterdensity.ThisdirectmeasurementmethodsignificantlysavestimeandresourcescomparingwithotherqPCRlibraryquantificationkitsonthemarket.ThisisachievedbyprovidingaconstructedlibraryratherthanasingleDNAfragmentasacontrolstandardwithknownclustergenerationdensityonIllumina^®sequencingplatforms.  

TheMCNext™SYBR^®FastqPCRLibraryQuantificationKitutilizesMCLAB’shighperformancefastPCRenzymebyaPhiXgenomebasedLibraryStandards(forseven10-folddilutions)withanaveragesizeof570bp,providingquickandaccuratequantificationwith0.0001–100pMdynamicrangein60min.Basedonthequantification,theresultcouldbedirectlyconvertedtoclusternumbersforloadingvolumereferences.

TheMCNext™SYBR^®FastqPCRLibraryQuantificationKitisarapidsolutionforyourlibraryconstructionQCtoaddvaluetoyourworkflowandincreaseconfidenceinyourresults.TheproprietaryPhiXLibraryStandardsconsistofagroupofIllumina^®sequencing adapter-ligatedDNAfragmentsderivedfromthePhiXgenomewithwell-characterizedsequenceandwell-balancedGCcontent(50%).TousethislibraryasacontrolforlibrariesconstructedforsequencingonIllumina^®sequencingplatformsisfarsuperiorthanasingleDNAfragmentascontrolstandardusedbyotherqPCRquantificationkitsonthecurrentmarket.Itcanberapidlyinterpolatedfortheestimatingclusterdensityandservesasaquantificationbaselineforsamplelibrarieswithouttheneedtohaveanothersequencingrunforaclusterdensitymeasurementpurpose.Itisanexcellentcontrolwithmeasuredclustergenerationconversionparametertoexaminethelibraryqualityandquantity,allowsyoutoquicklydetermineifanerrorisrelatedtothesamplepreparationbeforeahigh-costsequencingrun.Quantificationisfast,directandaccuratebyextrapolationagainstthestandardcurvegeneratedusingtheseven10-foldLibraryStandardsdilutions,followedbyasimpleclusternumberconversion.

TheMCNext™SYBR^®FastqPCRLibraryQuantificationKitisavailablein3types(RegularROX,LowROXandUniversal)basedontheinternalreferencedyepreferencesofthePCRThermocyclers.Thepackagesincludeready-to-use2XMasterMix,10XPrimerMixandPhiXLibraryStandards.

Features

TheMCNext™SYBR^®FastqPCRLibraryQuantificationKitprovidesresearcherswithanaccurateandsensitivemethodforquantifyingNGSlibraries.

• Directquantifylibraryclusterdensitybeforeloadingthesamples
• AccuratelibraryquantificationusingPhiXgenomebasedLibraryStandards
• Consistentlibraryquantificationacrossabroadrangeofsamples
• HighperformanceandfastPCRenzyme,highsensitivityandrapiddetection,simplifiedsteps
• Adaptabletohigh-throughputlibraryquantificationforlargerbarcodingapplications
• TailoredforlibrariesconstructedbyIllumina^®TrueSeqandNexteraworkflows

ConvenientpackagesforalltypesofrealtimePCRplatforms