Description:
OneTaq®HotStartDNAPolymeraseisanoptimizedblendofTaqandDeepVentR™DNApolymerasescombinedwithanaptamer-basedinhibitor.The3´→5´exonucleaseactivityofDeepVentDNAPolymeraseincreasesthefidelityandrobustamplificationofTaqDNAPolymerase(1)andthehotstartformulationcombinesconveniencewithdecreasedinterferencefromprimerdimersandsecondaryproducts.TheOneTaqReactionBuffersandHighGCEnhancerhavebeenformulatedforrobustyieldswithminimaloptimization,regardlessofatemplate’sGCcontent.
TheinhibitorbindsreversIBLy,blockingpolymeraseactivityattemperaturesbelow45°C.Thepolymeraseisactivatedduringnormalcyclingconditions,allowingreactionstobesetupatroomtemperature.OneTaqHotStartDNAPolymerasedoesnotrequireaseparatehightemperatureincubationsteptoactivatetheenzymeandcanbeusedintypicalTaq-basedcyclingprotocols.OneTaqHotStartDNAPolymeraseissuppliedwithtwo5Xbuffers:(StandardandGC),aswellasaHighGCEnhancersolution.Formostroutineand/orAT-richamplicons(LamBDa,etc.)orcomplexampliconswithupto~65%GCcontent,OneTaqStandardReactionBufferprovidesrobustamplification.ForGC-richamplicons,theOneTaqGCReactionBuffercanimprovebothperformanceandyield.ForparticularlyhighGCordifficultamplicons(i.e.≥75%GC),theOneTaqHighGCEnhancercanbeaddedatafinalconcentrationof10–20%toreactionscontainingOneTaqGCReactionBuffer.
Highlights
ProductSource
AnE.colistrainthatcarriestheTaqDNAPolymerasegenefromThermusaquaticusYT-1andanE.colistrainthatcarriestheDeepVentRDNAPolymerasegenefromPyrococcusspeciesGB-D.
ReagentsSupplied
Thefollowingreagentsaresuppliedwiththisproduct:
| Storeat(°C) | Concentration |
OneTaq®StandardReactionBuffer | -20 | 5X |
OneTaq® GCReactionBuffer | -20 | 5X |
OneTaq HighGCEnhancer | | 50% |
Notes:
TheOneTaqHighGCEnhancershouldnotbeusedalone.ItshouldbeaddedonlytotheOneTaqGCReactionBufferandwilltypicallyenhanceyieldswhenotherconditionshavefailed.ProductspecificationsforindividualcomponentsintheOneTaq HotStartDNAPolymerasemixareavailableseparately.