咨询热线：4000-520-616TheUltraIIDNALibraryPrepKitforIlluminameetsthechallengeofconstructinghighqualitylibrariesfromever-decreasinginputquantities. Thereagentsforeachstepinthelibrarypreparationworkflowhavebeenreformulatedtoenablehighyieldpreparationofhighqualitylibrariesfrom500picogramsto1microgramofinputDNA. ThisnewgenerationofNEBNextreagentsusesafast,streamlined,automatableworkflowandenablesuseoffewerPCRcycleswhilealsoimprovingGCcoverage.ThekitisalsocompatIBLewithPCR-freeworkflowsandiseffectivewithchallengingsamplessuchasFFPEDNA.Nowalsoavailablewith optionalSPRIselect®beads forclean-upandsize-selectionsteps.Advantages

Getmoreofwhatyouneed,withthehighestlibrary yields

Usetogenerate highqualitylibraries evenwhenyouhaveonlylimitedamountsofDNA,withinputsaslowas500pg

PreparelibrariesfromALLofyoursamples,including GC-richtargets and FFPEDNAsamples

Improveyieldsandqualityfor targetenrichment applications

Savetimewith streamlinedworkflows ,reducedhands-ontimeandautomationcompatibility,andenjoytheflexibilityofkitormoduleformatsTolearnmoreabouthowNEBNextUltraIIaddresseslowerinputamountsandchallengingsampletypes,downloadour technicalnote.

NEBNextUltraIIproducesthehighestlibraryyieldsandconversionratesfromabroadrangeofinputamountsA: LibrarieswerepreparedfromHumanNA19240genomicDNAusingtheinputamountsandnumbersofPCRcyclesshown.Manufacturers’recommendationswerefollowed,withtheexceptionthatsizeselectionwasomitted.B: LibrarieswerepreparedfromHumanNA19240genomicDNAusingtheinputamountsandlibraryprepkitsshownwithoutanamplificationstep,followingmanufacturers\'recommendations.qPCRwasusedtoquantitateadaptor-ligatedmolecules,andquantitationvalueswerethennormalizedtotheconversionrateforUltraII.TheUltraIIKitproducesthehighestrateofconversiontoadaptor-ligatedmolecules,forabroadrangeofinputamounts.Viewadditionaldataonlibraryyield NEBNextUltraIIprovidesuniformGCcoverageformicrobialgenomicDNAoverabroadrangeofGCcompositionandinputamountsLibrariesweremadeusing500pg,1ngand100ngofthegenomicDNAsshownandtheUltraIIDNALibraryPrepKitandsequencedonanIlluminaMiSeq^®.ReadsweremappedusingBowtie2.2.4andGCcoverageinformationwascalculatedusingPicard\'sCollectGCBiasMetrics(v1.117).Expectednormalizedcoverageof1.0isindicatedbythehorizontalgreyline,thenumberof100bpregionsateachGC%isindicatedbytheverticalgreybars,andthecoloredlinesrepresentthenormalizedcoverageforeachlibrary.Viewadditionaldataonlibraryquality NEBNextUltraIIprovidessuperioryieldsinPCR-freeworkflowsLibrariesweregeneratedfrom100ngofHumanNA19240genomicDNAusingthelibraryprepkitsshown,followingmanufacturers\'recommendations,andwithnoamplificationstep.LibraryyieldsweredeterminedbyqPCRusingtheNEBNextLibraryQuantKitforIllumina.TheNEBNextUltraIIKitproducesthehighestyields.

NEBNextUltraIIlibrariesprovidethehighestqualitysequencedatainPCR-freeworkflows

PCR-freelibrariesweregeneratedfrom100ngofHumanNA19240genomicDNAusinglibraryprepkitsshown,followingmanufacturers’recommendations,andwithnoamplificationstep.LibrariesweresequencedontheIlluminaNextSeq500.ReadsweremappedtotheGRCh37referenceusingBowtie2.2.4. ThisdataillustratesthatNEBNextUltraIIDNALibraryPrepKitenableshighqualitysequencedatainPCR-freeworkflows,evenwithlowinputamounts.%Mapped: ThepercentageofreadsmappedtohumanGRCh37reference. %Duplication: Thepercentageofmappedsequencethatismarkedasduplicate. %Chimeras: Thepercentageofreadsthatmapoutsideofamaximuminsertsizeorthathavethetwoendsmappingtodifferentchromosomes. NEBNextUltraIIenablesconstructionofhighqualitylibrariesfromFFPEDNAsamples LIBRARYKITTOTALREADS%DUPLICATION%MAPPED%CHIMERAS

UltraII	3,685,029	0.02	97.00	1.10
KapaHyper	3,679,136	0.02	97.00	2.00
TruSeqPCR-Free	3,681,643	0.05	96.65	1.38

Librarieswerepreparedfrom17–30ngofhumanDNAextractedfromtheFFPEtissuesampleslisted,amplifiedusing10cyclesofPCRandsequencedontheIlluminaMiSeq. ReadsweremappedtotheGRCh37referencegenomeusingBowtie2.2.4.%Mapped: ThepercentageofreadsmappedtoHumanGRCh37reference. %MappedinPairs: Thepercentageofreadswhosematepairwasalsoalignedtothereference.%Duplication: Thepercentageofmappedsequencethatismarkedasduplicate. %Chimeras: Thepercentageofreadsthatmapoutsideofamaximuminsertsizeorthathavethetwoendsmappingtodifferentchromosomes.ThisdataillustratesthattheNEBNextUltraIIDNALibraryPrepKitforIlluminaenableshighqualitysequencedata,evenwithlowinputamountsofFFPEDNA. ViewadditionaldatafromFFPEDNAsamples

NEBNextUltraIIprovidessuperioryieldsfortargetenrichmentapplications.ForNimblegen^® enrichment,librarieswerepreparedwith100ngofHumanHG02922genomicDNA,usingNEBNextUltraIIwiththeNEBNextAdaptorandindexprimers,ortheKapaLibraryPreparationKitIlluminaPlatformswiththeSeqCap^® AdapterKit(Roche).TargetenrichmentwasperformedwithNimbleGenSeqCapHumanExomev3.ForSureSelect^XT enrichment,librarieswerepreparedwith200ngofHumanHG02922genomicDNA,usingNEBNextUltraIIorKapaHyperwiththeNEBNextAdaptorandindexprimers,ortheSureSelect^XT ReagentKit,Illumina(ILM)platformswithHerculase^® IIFusionDNAPolymerase.TargetenrichmentwasperformedwithSureSelect^® HumanAllExonV6.Manufacturers’recommendationswerefollowed,includinguseoftherecommendednumberofPCRcyclesforpre-andpost-capture.NEBNextUltraIIlibrariesresultedinthehighestpost-captureyieldsforbothenrichmentworkflows.Viewadditionaldataontargetenrichment

LotControl

Thelotsprovidedaremanagedseparatelyandqualifiedbyadditionalfunctionalvalidation.Individualreagentsundergostandardenzymeactivityandqualitycontrolassays,andalsomeetstringentcriteriaintheadditionalqualitycontrolslistedoneachindividualcomponentpage

ReagentsSupplied

Thefollowingreagentsaresuppliedwiththisproduct:

FFPEDNADNAINPUT(ng)LIBRARYYIELDSINng%MAPPED%MAPPEDINPAIRS%DUPLICATION%CHIMERAS

KidneyTumor	17	132	91.5	96.1	0.48	3.0
LungTumor	20	232	90.1	94.9	0.42	4.1
LiverNormal	20	691	92.6	94.7	0.33	8.6
BreastTumor	30	514	91.9	95.1	0.37	4.5

	Storeat(°C)	Concentration
NEBNextUltraIIEndPrepEnzymeMix	-20
NEBNextUltraIIEndPrepReactionBuffer	-20
NEBNextLigationEnhancer	-20
NEBNextUltraIILigationMasterMix	-20
NEBNext®UltraIIQ5®MasterMix	-20	2X

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