Description:
T4DNAPolymerasecatalyzestheextensionofaprimedDNAtemplateinthe5'->3'direction.Thisenzymeexhibitsapowerful3'->5'exonucleaseactivity,whilelackinganyinherent5'->3'exonucleaseorstranddisplacementfunctions.
SpecificActivity:5,555U/mg
Application:
-3'-overhangremovaltoformbluntends
-5'-overhangfill-intoformbluntends
-Singlestranddeletionforsub-cloning
-Secondstrandsynthesisinsite-directedmutagenesis
-Probelabelingusingreplacementsynthesis
Source:
PurifiedfromastrainofE.colithatexpressestherecombinantT4DNAPolymerasegene.
Suppliedin:
100mMKPO4
1.0mMDTT
0.1mMEDTA
50%Glycerol
pH6.5@25°C
SuppliedWith:10xBlueBuffer
10xBlueBuffer:
500mMNaCl
100mMTris-HCl
100mMMgCl2
10mMDTT
pH7.9@25°C
UnitDefinition:
Oneunitisdefinedastheamountofenzymethatwillincorporate10nmolofdNTPsintoacid-precipitablematerialin30minutesat37°C.
RecommendedStorageCondition:-20ºC
Reference:
1.Tabor,S.andStruhl,K.(1989)InDNA-DependentDNAPolymerases.F.M.Ausebel,R.Brent,R.E.Kingston,D.D.Moore,J.G.Seidman,J.A.SmithandK.Struhl(Eds.),CurrentProtocolsinMolecularBiology,pp.3.5.10-3.5.12.2.Sambrook,J.,Fritsch,E.F.andManiatis,T.(1989)MolecularCloning:ALaboratoryManual,(2ndEd.),5.44-5.47.
2.Dale,R.,McClure,B.andHouchins,J.(1985)Plasmid,13,31-40.
3.Kunkel,T.A.,Roberts,J.D.andZakour,R.A.(1987)R.WuandL.Grossman(Eds.),MethodsEnzymol.,154,pp.367-382.SanDiego:AcademicPress.
4.Panet,A.,vandeSande,J.H.,Loewen,P.C.andKhorana,H.G.(1973)Biochemistry,12,5045-5050. |
