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ID:50167439
大小:294.14 KB
页数:10页
时间:2020-03-04
《Ion torrent建库试剂说明书.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、TechnicalDataSheetKAPALibraryPreparationKitKitCodesandComponentsIonTorrent™PlatformsKR0573-v1.13EndRepairBuffer(10X)62µlKK8300EndRepairEnzymeMix44µlKK8310Ligation&NickRepairBuffer(5X)123µlProductDescription8Ligation&NickRepairEnzyme53µlThisKAPALibraryPreparationKitisdesignedf
2、orlibrariesKAPAHiFiHotStartReadyMix(2X)*220µlthepreparationoflibrariesforsequencingontheIonLibraryAmplificationPrimerMix(10X)*44µlPersonalGenomeMachine™(PGM™)andIonProton™semiconductorsequencers.ThekitprovidesalloftheEndRepairBuffer(10X)413µlenzymesandreactionbuffersrequiredf
3、orconstructingKK8301EndRepairEnzymeMix295µllibrariesfrom100ng–1µgoffragmented,double-KK8311Ligation&NickRepairBuffer(5X)827µlstrandedDNAviathefollowingsteps:Ligation&NickRepairEnzyme354µl481.Endrepair:Produceblunt-ended,5'-phosphorylatedlibrariesKAPAHiFiHotStartReadyMix(2X)*1
4、480µlLibraryAmplificationPrimerMix(10X)*295µlfragments.2.Adapterligationandnickrepair:LigatedsDNA*KK8310andKK8311areavailableforPCR-freeworkflowsanddonotadapterstoblunt-endedlibraryfragmentsandcontainKAPAHiFiHotStartReadyMixorLibraryAmplificationPrimerperformnickrepairtoyield
5、libraryfragmentsflankedMix.byadapters.3.Libraryamplification(optional):PerformPCRtoQuickNotesamplifylibraryfragmentscarryingappropriateadaptersequencesonbothends.•Theprotocolprovidedhereisgeneric,andmayThekitprovidesalloftheenzymesandbuffersrequiredrequireadditionaltailoringa
6、ndoptimizationforlibraryconstructionandamplification,butdoesnotdependingontheintendedapplication.includeadaptersorSPRI®beads.Enzymeformulations•Separate,concentratedenzymeformulationsandandreactionbuffersforendrepair,andligationandnickreactionbuffersprovidethebestcombinationo
7、frepairaresuppliedinconvenient,concentratedformats.productstability,convenience,andefficiency.Adaptersareavailableseparately(KK8330,KK8331,KK8332orKK8333).Theseformulationsensuremaximum•Generousreagentvolumeshavebeenprovidedinstabilityofthereagentsandhighreactionefficiencies,
8、the48reactionkittoaccommodatedeadvolumeswhilesimplifyingreactionsetu
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