AThermostableDNA-andRNA-dependent(i.e.,reversetranscriptase)DNApolymeraseforbothPCRandRT-PCRapplications
FlexIBLe:UseasingleenzymeforbothPCRandRT-PCRapplications,greatlysimplifyingreactionoptimizationandsetup- Thermostable:EnablesamplificationofDNAandRNAtargetscontaininghighdegreesofsecondarystructure
MasterAmp™TthDNAPolymeraseisarecombinantDNAenzymefromThermusthermophilusthathasbothDNA-dependentandRNA-dependent(i.e.,reversetranscriptase)DNApolymeraseactivitiesupto~95°C.MasterAmpTthDNAPolymeraseishighlypurifiedusingaproprietaryprocedurethatvirtuallyeliminatescontaminatingbacterialDNA.
Applications
- PCRamplificationofDNAandone-stepRT-PCRofRNA.
- PCRamplificationofRNAandDNAtemplateshavingahighdegreeofsecondarystructure.
Optimizationof[Mg2+]and[Mn2+]:MagnesiumandmanganeseionconcentrationsshouldbeoptimizedforDNAsynthesis.TheoptimalconcentrationofeachmetalcationishighlydependentonthedNTPconcentrationandonthetemplate,primers,andprotocolused.Formanytemplates,theoptimalconcentrationofmagnesiumionsusingMasterAmpTthDNAPolymeraseisapproximately1.5mMiftheconcentrationofeachdNTPinthereactionis0.2mM.Ifusedwith1.5mMMg2+,theoptimalconcentrationofMn2+forRNA-dependentDNAsynthesisisapproximately0.5mMformanytemplates.ForanRT-PCRreaction,werecommend3.0mMMg2+and0.5mMMn2+formosttemplates.Alternatively,werecommendtheMasterAmp™PCROptimizationKitwithammoniumsulfateforrapidPCRoptimization.
UnitDefinition:OneunitofMasterAmpTthDNAPolymeraseconverts10nmolofdNTPsintoacid-insolublematerialin30minutesat70°Cunderstandardassayconditions.
StorageBuffer:50%glycerolcontaining50mMTris-HCl(pH7.5),100mMNaCl,0.1mMEDTA,0.5%Tween20,0.5%NP-40,and1mMDTT.
QualityControl:MasterAmpTthDNAPolymeraseistestedfortheabsenceofdetectablenonspecificexo-andendonucleaseandRNaseactivities,andarefunctionallytestedinPCR.
*PurchaseofMasterAmp™TthDNAPolymeraseincludesanimmunityfromsuitunderpatentsspecifiedintheproductinserttouseonlytheamountpurchasedforthepurchaser"sowninternalresearch.Nootherpatentrights(suchas5′NucleaseProcesspatentrights)areconveyedexpressly,byimplication,orbyestoppel.FurtherinformationonpurchasinglicensesmaybeobtainedbycontactingtheDirectorofLicensing,AppliedBiosystems,850LincolnCentreDrive,FosterCity,California94404,USA.Coveredbyissuedandpendingpatents.Forcompletelicensestatement,seep.iv.
**Coveredbyissuedand/orpendingpatents.
