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