Details
Source
Thermusaquaticus
Description
- TaqDNAPolymerase,Native isaThermostableenzymeofapproximately94kDafrom Thermusaquaticus
Applications
- ReplicatesDNAat74°Candexhibitsahalf-lifeof40minutesat95°C(1,2)
- CatalyzesthepolymerizationofnucleotidesintoduplexDNAinthe5"→3"directioninthepresenceofmagnesiumions
- Maintainsthe5"→3"exonucleaseactivity
- Lacksthe3"→5"exonucleaseactivity
- RecommendedforuseinPCRandprimerextensionreactionsatelevatedtemperaturestoobtainawiderangeofDNAproducts upto10Kb
- RecommendedforuseinspecialPCRapplications,wheretracesof E.coli genomicDNAmayinterferewithamplificationspecificity
ReagentsSupplied
10XTaqBufferA
10XTaqBufferB
10XTaqBufferC
UnitDefinition
Oneunitisdefinedastheamountofenzymerequiredtocatalyzetheincorporationof10nmolesofdNTPintoacid-insolublematerialin30minutesat70°C
10XReactionBuffer
10XTaqBufferA(optimizationbufferwithoutMgCl2):AllowstooptimizeMgCl2 concentration
10XTaqBufferB(generalapplication,upto10kb): Contains15mM MgCl2 andis optimizedforusewith0.2 mMofeachdNTP
10XTaqBufferC(colored): EnrichedwithtwogeltrackingdyesandagelloADIngreagent,enablesdirectloadingofPCRproductsontoanagarosegel
StorageBuffer
20mMTris-HCl(pH8.0at22°C)
100mMKCl
0.1mMEDTA
1mMdithiothreitol
50%glycerol
StABIlizers
AssayConditions
25mMTris-HCl(pH9.5at25°C)
50mMKCl
10mMMgCl2
1mMdithiothreitol
200μMeachofdCTP,dGTP,dTTP,anddATP(amixofunlabeledand[α-32P]dATP)
10μgactivatedcalfthymusDNA
1mg/mlbovineserumalbumin
15 µgactivatedcalfthymusDNA
Totalreactionvolumeis50 µl
QualityControl
Allpreparationsareassayedforcontaminatingendonuclease,3"-exonuclease,andnonspecificsingle-anddouble-strandedDNaseactivities.Typicalpreparationsaregreaterthan95%pure,asjudgedbySDSpolyacrylamidegelelectrophoresis.
StorageConditions
Storeat-20°C
Shippedondryice
Downloads
MSDSPDF
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
