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NEB//E6627S/8 reactions
  • NEB//E6627S/8 reactions

NEB//E6627S/8 reactions

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    • Description:

      Sample

      ?
      ?
      TheNEBNextDirectBRCA1/BRCA2PanelenrichesthecompleteexoncontentofBRCA1andBRCA2genesfornext-generationse-quencinganalysis.NEBNextDirectemploysauniquehybridization-basedenrichmentworkflowthathybridizesbaitsdirectlytogenomicDNA,withouttheneedforupfrontlibrarypreparation.TheBRCA1/BRCA2paneldemonstratesextremelyhighspecificityandunmatchedcoverageuniformityacrossawiderangeofDNAinputs,allowinghighlysensitivecallingofgermlineandsomaticvariantswhilemaximizingsequencerefficiency.

      Benefits:

      • Generatefull(100%)coverageofallproteincodingregionsinBRCA1andBRCA2genes
      • Obtainhighlyuniformsequencingacrossexontargets-100%ofbasepairsinpanelhavecoveragegreaterthan20%ofthemeantargetcoverage.
      • Maximizeefficiencywitha1-dayworkflowthatcombineshighlyspecificenrichmentwithlibrarypreparation
      • ProducehighdepthsoftargetcoverageacrossawiderangeofDNAinputamountsforgermlineandsomaticvariantcalling.

      Figure1.NEBNextDirectWorkflow.
      fast hybridization workflow

      TheNEBNextDirectBRCA1/BRCA2PaneldelivershighlyefficientenrichmentofBRCA1andBRCA2codingregionswithahighpercentageofreadsmappingtotargets
      Thishistogramshowsthepercentofreadsalignedtothehumangenome,andthepercentofreadsmappedtothetargetsincludedintheBRCA1/BRCA2PanelacrossdifferentinputDNAamounts.10ng,100ngand1μgofpurifiedgenomicDNAwasenrichedusingtheNEBNextDirectBRCA1/BRCA2Panel.Sequencingreadsweregeneratedonanllumina?MiSeq?with2x75bpreads,8bpSampleIDand12bpuniquemolecularidentifier.SequencingreadalignmentswereperformedwithBWA-MEM,andPCRduplicateswerefilteredusingtheUMIs.


      TheNEBNextDirectBRCA1/BRCA2Panelproduceshighlyuniformcoverageacrosstargetregions,maximizingsequencerefficiency
      Thishistogramshowsthenormalizedmeancoverageacrosseachofthe49exontargetsoftheBRCA1andBRCA2genes.50ngofDNAwasenrichedusingthe17kbNEBNextDirectBRCA1/BRCA2Panel.SequencingreadsweregeneratedonanIlluminaMiSeqwith2x75bpreads,an8bpsampleIDand12bpuniquemolecularidentifier.SequencingreadalignmentswereperformedwithBWA-MEM,andPCRduplicateswerefilteredusingtheUMIs.


      TheNEBNextDirectBRCA1/BRCA2PanelcanbeusedtoprovidedepthsofsequencingcompatIBLewithbothgermlineandsomaticvariantcalling
      Thisfiguredemonstratesde-duplicatedcoverageofBRCA1andBRCA2genesasafunctionofsequencingdepthacrossarangeofinputDNAamounts.PurifiedgenomicDNAwasenrichedusingtheNEBNextDirectBRCA1/BRCA2Panel.SequencingreadsweregeneratedonanIlluminaMiSeqwith2x75bpreads,the8bpsampleIDand12bpuniquemolecularidentifier.SequencingreadalignmentswereperformedwithBWA-MEM,andPCRduplicateswerefilteredusingtheUMIs.


      TheNEBNextDirectBRCA1/BRCA2Panelproducesevendepthsofcoverageacrosstargetregions

      IntegrativeGenomeViewer(IGV)plotshowingthereadcoverageacrossexon1oftheBRCA1gene.50ngofDNAwasenrichedusingthe17kbNEBNextDirectBRCA1/BRCA2Panel.SequencingreadsweregeneratedonanIlluminaMiSeqwith2x75bpreads,an8bpsampleID,and12bpuniquemolecularidentifier.SequencingreadalignmentswereperformedwithBWA-MEM,andPCRduplicateswerefilteredusingtheUMIs.

      KitComponents

      Thefollowingreagentsaresuppliedwiththisproduct:

      Storeat(°C)Concentration
      NEBNextDirect?HybridizationAdditive-20
      NEBNextDirect?3′BluntingEnzymeMix-20
      NEBNextDirect?dA-TailingEnzyme-20
      NEBNextDirect?3′Adaptor-20
      NEBNextDirect?Ligase-20
      NEBNextDirect?5′BluntingEnzymeMix-20
      NEBNextDirect?5′UMIAdaptor-20
      NEBNextDirect?CleavingEnzymeMix-20
      NEBNextDirect?FFPEPhosphorylationEnzyme-20
      NEBNextDirect?BeadWash125
      NEBNextSamplePurificationBeads25
      NEBNextDirect?StreptavidinBeads4
      NEBNextDirect?HybridizationWash(HW)4
      NEBNextDirect?BeadPrepBuffer4
      NEBNextDirect?HybridizationBuffer4
      NEBNextDirect?dA-TailingBuffer4
      NEBNextDirect?AdaptorLigationBuffer4
      NEBNextDirect?5′BluntingBuffer4
      NEBNextDirect?CleavingBuffer4
      NEBNextDirect?BeadWash24
      NEBNextDirect?Q5MasterMix-20
      NEBNextDirectBRCA1/BRCA2Baits-20
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