Recombinant Human Turbo Dicer Enzyme Kit PDF b2bd3b86
Recombinant Human Turbo Dicer Enzyme Kit
GenlantisAdivisionofGeneTherapySystemsIncPRODUCTSUMMARYCatNo152000250UnitsKitComponentsContent_RecombinantTurboDicerEnzyme1tubex100ul05unitulDicerReactionBuffer1tubex50ul10mMATP1tubex50ulDicerStopSolution1tubex100ul5XBSA1tubex50ulNucleasefreeWater1tubex1mlShippingandStorageTheRecombinantHumanTurboDicerEnzymeKitisshippedfrozenFormaximumstabilityandlongtermuseimmediatelystoreat20CuponreceiptAllcomponentsarestableforsixmonthswhenstoredproperly50mMMgClSolution1tubex100ulDescriptionTheRecombinantHumanTurboDicerEnzymeKitusesanultraactiveformofhumanrecombinantdicerenzymethatcancleavemorethan95ofdsRNAtemplateinto22bpsiRNAswithin2hoursunderoptimizedreactionconditionsFigure1ThisprovidessufficientsiRNAsineverysinglespeciestoachieveeffectivegenesilencingFig1EfficientDigestionofDoubleStrandedRNAwithRecombinantHumanTurboDicerEnzymeI3AgaroseGelLaneSample1100bpmarker2lug700bpdsRNA31ug700bpdsRNA15hourdigestionwith1unitDicerEnzyme41ug700bpdsRNA2hourdigestionby1unitTurboDicerEnzymeRecombinantHumanTurboDicerEnzymeKitPage1of2RecombinantHumanTurboDicerEnzymeKitNOTETheRecombinantHumanDicerEnzymeKitisintendedFORRESEARCHUSEONLYMETHODSANDPROCEDURESPreliminaryNotes1WerecommendthatyouusedsRNAtemplatesof5001000bpinlengthThedicerenzymemightnotdigestwellifthedsRNAissmallerthan300bp2ForoptimalresultswerecommendusingtheGTSTurboScriptT7RNATranscriptionKitCatalogT510003togeneratedoublestrandedRNAdsRNAdsRNAsgeneratedwithTurboScriptcanbeuseddirectlywiththeRecombinantTurboDicerEnzymeKitwithoutpurificationhoweverdsRNAspurifiedusingstandardlithiumchlorideorequivalentprocedurescangiveslightlybetterresultsGenerationofsiRNAUsingRecombinantTurboDicerEnzyme1KeeptheDicerReactionBufferatroomtemperaturewhileassemblingthereactionThefollowingamountsareforasingle10ulreaction3ulxNucleasefreewaterxuldsRNA1ug1ul10mMATP1ul5XBSA2ul50mMMgCl1ul10XDicerReactionBuffer2ulRecombinantTurboDicerEnzyme05unitulNOTEJUgofdsRNAcontroltemplatewillyieldabout05ugofdicedsiRNAdsiRNAafterpurificationwithGTSRNAPurificationColumns1amp2thisissufficientforoneortwotransfectionsin24wellplatesusingtheGeneSilencersiRNATransfectionReagentFormoredsIRNAsscaleupthereactionvolumeproportionallyIMPORTANTAvoidusingexcessRecombinantDicerEnzymeasitmaydecreasetheamountofdsiRNAthatcanbegeneratedfromthedigestio
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