Description:


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TheNEBNext??Ultra?DNALibraryPrepKitforIlluminacontainsenzymesandbuffersthatareidealtoconvertasmallamountofDNAinputintoindexedlibrariesfornext-generationsequencingontheIlluminaplatform(Illumina,Inc).TheworkflowofNEBNextUltraDNALibraryPrepKitforIlluminaisveryuser-friendlyandfastwithminimalhands-ontime.Eachofthesecomponentsmustpassrigorousqualitycontrolstandardsandarelotcontrolled,bothindividuallyandasasetofreagents.
UltraDNALibraryPreparationWorkflowforIllumina
NEBNextUltra?DNAprovideshighlibraryyieldsevenwithlow?inputsanddifficultsamples
Alllibrarieswerepreparedwith5ngofinputDNAandrunonanAgilentBioanalyzer?.
A.E.coligenomicDNAlibraries.1:Ladder;2:LibrarypreparedusingNEBNextDNA?LibraryPrepMasterMixSet(E6040);3:LibrarypreparedusingNEBNextUltraDNA?LibraryPrepKit(E7370).With5nginputs,libraryyieldsaresignificantlyhigherusing?theNEBNextUltra?DNAKit.
B.FFPEsample.HumanlungtumorgenomicDNAfroma66-yearoldmale,extractedfrom?a10-yearoldFFPEblock(Biochain,Inc.).1:LibrarypreparedusingNEBNextUltraDNA?LibraryPrepKit(E7370).2:InputDNA(highlydegradedFFPEgenomicDNA).Readdepthcorrelationshowsconsistenthighcoveragefor5ng-1μginputamounts
LibrarieswerepreparedwithvariousamountsofhumanIMR90gDNA(5,50,and1000ng)andPCRamplified.APCR-freelibrarywasalsopreparedwith1000nghumanIMR90gDNA.Sequencecoverageof10kbwindowsofthehumangenomewasanalyzedonGalaxy,andPearsoncorrelationcoefficient(R2values)werecalculatedusingMicrosoft?Excel?.Alllibrariesarehighlycorrelatedwitheachother,independentofDNAinputaswellasPCRamplification.HighcoveragewasachievedforallDNAinputamounts.
NEBNextUltraDNAKitsdisplaystrongperformancewithinputamountsaslowas5ng

InputDNA:IMR90genomicDNA(humanfibroblast).
PFMismatchRate:Therateofbasesmismatchingthereferenceforallbasesalignedtothereferencesequence.
PercentDuplication:Thepercentageofmappedsequencethatismarkedasduplicate.
PercentChimeras:Thepercentageofreadsthatmapoutsideofamaximuminsertsizeorthathavethetwoendsmappingtodifferentchromosomes.
ThisdataillustratesthestrongperformanceoftheNEBNextUltraDNALibraryPrepKitforIllumina,evenwith5ngofhumangenomicDNAinput.Mismatchrate,%duplicationand%chimerasarealllow,andsimilar,regardlessofinputamount.
FunctionalValidation
EachsetofreagentsisfunctionallyvalidatedtogetherthroughconstructionandsequencingofaDNAlibraryontheIlluminasequencingplatform.LotControl
Thelotsprovidedaremanagedseparatelyandqualifiedbyadditionalfunctionalvalidation.Individualreagentsundergostandardenzymeactivityandqualitycontrolassays,andalsomeetstringentcriteriaintheadditionalqualitycontrolslistedoneachindividualcomponentpage
ReagentsSupplied
Thefollowingreagentsaresuppliedwiththisproduct:
| Storeat(°C) | Concentration |
NEBNextEndPrepEnzymeMix | -20 | |
NEBNextEndRepairReactionBuffer | -20 | 10X |
Blunt/TALigaseMasterMix | -20 | |
NEBNext?LigationEnhancer | -20 | |
NEBNext®Q5®HotStartHiFiPCRMasterMix | -20 | |