Details
Source
BacteriophageT4ofEscherichiacoli
Description
- T4DNAPolymeraseexhibits5"→3"polymeraseand3"→5"exonucleaseactivities(1,2)
- Requiresthepresenceofasingle-strandedDNAtemplateandaprimer
- Exonuclease,strongerthanthatfoundinDNAPolymeraseI,ismoreactiveonsingle-strandedDNAthanondouble-strandedDNA
- Ultrapurerecombinantenzyme
Applications
- Addslabelednucleotidestotherecessed3"-endsofDNAfragments
Exonucleaseactivitycanbeusedtoremoveoneorafewnucleotidesfrom3’-endofdouble–strandedDNA - T4DNAPolymeraseenzymesuitablefor:
-3’overhangremovaltoformbluntends(3,4)
-5’fill-intoformbluntends(3,4)
-Probelabelingusingreplacementsynthesis(3,4)
-Singlestranddeletionsubcloning(5)
-Secondstrandsynthesisinsite-directedmutagenesis(6)
ReagentsSupplied
10XT4DNAPolymeraseReactionBuffer
UnitDefinition
Oneunitistheamountofenzymerequiredtoincorporate10nmolesoftotalnucleotideintoacidinsolubleformfor30minutes at37°C
Concentration
5-10Units/µl
AssayConditions
67mMpotassiumphosphate(pH8.8)
6.7mMMgCl2
1mMdithiothreitol
16.6mMammoniumsulfate
6.7µMEDTA
167µg/mlbovineserumalbumin
0.033mM[α-32P]dATP
StorageConditions
Storeat-20ºC
Productshippedondryice
Downloads
CertificateofAnalysisPDF -CurrentLot
MSDSPDF
References
1.Goulian,M.,Lucas,Z.J.andKornberg,A.(1968)J.Biol.Chem.243,627-638
2.Lehman,I.R.(1981)Enzymes14,51-65
3.Tabor,S.andStruhl,K.(1989)CurrentProt.inMol.Bio.(Ausubel,F.M.,etal.,eds)pp.3.5.10-3.5.12
4.Sambrook,J.,Fritsch,E.F.andManiatis,T.(1989)Mol.Cloning:ALabManual,SecondEd.pp.5.44-5.47
5.Dale,R.,McClure,B.andHouchins,J.(1985)Plasmid13,31-40
6.Kunkel,T.A.,Roberts,J.D.andZakour,R.A.(1987)MethodsEnzymol.154,367-382
