Description: Choo-ChooCloningTMKitsarehighlyefficientdirectionalPCRcloningkitsforrapid,ligaseandrestrictionenzyme-independentcloningofPCRproducts.TheyallowtocloneanyPCRfragmentintoanylinearizedvectoratanylocation.Bysimplyincubatingonice,theendsofaPCR-generatedDNAfragmentcanbepreciselyfusedintoaDNAvectorwith6bp(ormore)overlappinghomologoussequence.Thesystemisveryrobust.Upto8PCR-generatedDNAfragmentscanbeassembledandclonedintoonepiece(upto10kbinonestep).Thesystemishighlyefficient,with98-100%positivecolonies. 
Function: ThefunctionofChoo-ChooCloningTMKitsdependonMCLAB'sproprietaryenzymesystems.Thereisnoneedforrestrictionenzymedigestion,ligation,orblunt-endpolishing.Anyextraunwantedbasescanbeeliminated inthefinalconstruct.ThelinearizedvectorcanbegeneratedbyPCRorrestrictionenzymedigestion.ThePCRfragmentscanbegeneratedbyTaqDNApolymeraseorotherhighfidelityDNApolymerases.TheadditionofaAbyTaqDNApolymeraseisnotrequiredandhasnoeffectoncloningefficiency.IfthePCRproductisamplifiedfromaplasmidtemplate,thenitneedstobegel-purifiedtoreducebackground.InadditiontoPCRcloning,Choo-ChooCloningTMKitsalsohavethefollowingaplications:adaptor,linkerandtagadditionbeforeandaftertheinserts,andgenesynthesis. Features: -Cloneanyinsertintoanyvectoryouchooseatanylocation -Norestrictiondigestion,phosphatasetreatment,orligationrequired -Multiplefragments(upto8pieces) -BroadPCRsizes(upto10kb) -Finalconstructsareseamlesswithnoextraorunwantedbasepairs -Simple45minutesingle-tubereactiononiceprotocol -Highefficiencywith>=98%positivecolonies -Multiplefunctions:compatIBLewithadaptor,linkerandtagadditionbeforeoraftertheinserts;compatiblewithgenesynthesis -Highthroughputapplication RecommendedStorageConditions: StoreBox1at-20°C. StoreBox2at-80°C. Components: TheChoo-ChooCloningTMKitsareavailablein10-,20-,100-,and96-reactionsizes. Cat# | Box1 | Box2 | | 10xChoo-ChooCloningTM ReactionBuffer | Choo-ChooCloningTM EnzymeMix | DH5a Competent E.coli cells | CCK-10(10rxn) | 20µl | 20µl | 50µlx10tubes | CCK-20(20rxn) | 40µl | 40µl | 50µlx20tubes | CCK-100(100rxn) | 200µl | 200µl | 50µlx100tubes | CCK-096(96rxn) | 96X5µl | 96x50μl(96-wellplate) | CCK-384(384rxn) | 384X5µl | 8x2.5ml |
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