Ion torrent建库试剂说明书.pdf

《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