NEBE7645S/NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®/现货

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TheUltraIIDNALibraryPrepKitforIlluminameetsthechallengeofconstructinghighqualitylibrariesfromever-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.

NEBNextUltraIIproducesthehighestlibraryyieldsandconversionratesfromabroadrangeofinputamounts

A: LibrarieswerepreparedfromHumanNA19240genomicDNAusingtheinputamountsandnumbersofPCRcyclesshown.Manufacturers’recommendationswerefollowed,withtheexceptionthatsizeselectionwasomitted.

B: LibrarieswerepreparedfromHumanNA19240genomicDNAusingtheinputamountsandlibraryprepkitsshownwithoutanamplificationstep,followingmanufacturers'recommendations.qPCRwasusedtoquantitateadaptor-ligatedmolecules,andquantitationvalueswerethennormalizedtotheconversionrateforUltraII.TheUltraIIKitproducesthehighestrateofconversiontoadaptor-ligatedmolecules,forabroadrangeofinputamounts.Viewadditionaldataonlibraryyield
 NEBNextUltraIIprovidesuniformGCcoverageformicrobialgenomicDNAoverabroadrangeofGCcompositionandinputamounts

Librariesweremadeusing500pg,1ngand100ngofthegenomicDNAsshownandtheUltraIIDNALibraryPrepKitandsequencedonanIlluminaMiSeq^®.ReadsweremappedusingBowtie2.2.4andGCcoverageinformationwascalculatedusingPicard'sCollectGCBiasMetrics(v1.117).Expectednormalizedcoverageof1.0isindicatedbythehorizontalgreyline,thenumberof100bpregionsateachGC%isindicatedbytheverticalgreybars,andthecoloredlinesrepresentthenormalizedcoverageforeachlibrary.Viewadditionaldataonlibraryquality
 NEBNextUltraIIprovidessuperioryieldsinPCR-freeworkflows

Librariesweregeneratedfrom100ngofHumanNA19240genomicDNAusingthelibraryprepkitsshown,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
 

LIBRARYKIT	TOTALREADS	%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:

FFPEDNA	DNAINPUT(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