Description

• Thermostableenzymeofapproximately94kDafrom Thermusaquaticus
• Ultrapure,Recombinantprotein

Applications

• RecommendedforuseinPCRandprimerextensionreactionsatelevatedtemperaturestoobtainawiderangeofDNAproducts upto10Kb
• ReplicatesDNAat74°Candexhibitsahalf-lifeof40minutesat95°C(1,2)
• CatalyzesthepolymerizationofnucleotidesintoduplexDNAinthe5"→3"directioninthepresenceofmagnesiumions
• Maintainsthe5"→3"exonucleaseactivity
• Lacksthe3"→5"exonucleaseactivity
• AddsextraAatthe3"-ends

Advantages

• Helpsvisualizetheadditionofthepolymerasetothereaction
• Confirmscompletemixing
• EnablesdirectloADIngofPCRproductsontoanagarosegelwithouttheadditionofagelloadingbuffer
• Addeddyesallowtrackingofelectrophoresisprogress
• DoesnotinterferewithPCRperformance
• Doesnotinterferewithmostdownstreamapplications

Note:Notrecommendedforanyapplicationsusingabsorbanceorfluorescenceexcitation

ReagentsSupplied

10XTaqBufferA
10XTaqBufferB

UnitDefinition

Oneunitisdefinedastheamountofenzymerequiredtocatalyzetheincorporationof10nmolesofdNTPintoacid-insolublematerialat74°Cin30minutes

ReactionBuffer

10xTaqBufferA(optimizationbufferwithoutMgCl₂): allowstooptimizeMgCl₂ concentration
10xTaqBufferB(generalapplication,upto10kb): contains15mM MgCl₂ andis optimizedforusewith0.2 mMofeachdNTP

StorageBuffer

20mMTris-HCl(pH8.0at22°C
100mMKCl
0.1mMEDTA
1mMdithiothreitol
50%(v/v)glycerol
Stablizers 

AssayConditions

50mMTris-HCl(pH9.0at25°C)
50mMNaCl
50mMMgCl₂ 
200µMeachofdATP,dCTP,dGTPanddTTP(amixofunlabeledand[³H]dTTP
10µgactivatedcalfthymusDNA
0.1mg/mlbovineserumalbumin
Finalvolumeof50µl

QualityControl

Allpreparationsareassayedforcontaminatingendonuclease,3"-exonuclease,andnonspecificsingle-anddouble-strandedDNaseactivities
Typicalpreparationsaregreaterthan95%pure,asjudgedbySDSpolyacrylamidegelelectrophoresis

StorageConditions

Storeat-20°C
Productshippedondryice

References 

1.Chien,A.,Edgar,D.B.andTrela,J.M.(1976)J.Bacteriol.127,1550 
2.Kaledin,A.S.,Sliusarenko,A.G.andGorodetskii,S.I.(1980)Biokhimiya45,644