Description | T4DNAPolymerasecatalyzestheextensionofaprimedDNAtemplate(inthepresenceoftemplateandprimer)inthe5′→3′direction.Thisenzymeexhibitsapowerful3′→5′exonucleaseactivitywhichismuchmoreactivethanthatfoundinDNAPolymeraseI(E.coli),whilelackinganyinherent5′→3′exonucleaseorstranddisplacementfunctions. |
Applications | - BluntingofDNAends:fill-inof5'-overhangsor/andremovalof3'-overhangs
- BluntingofPCRproductswith3'-dAoverhangs
- Singlestranddeletionsubcloning(Ligation-independentcloningofPCRproducts).
- Secondstrandsynthesisinsite-directedmutagenesis.
- Probelabelingusingreplacementsynthesis.
|
Source | ArecombinantE.colistraincarryingaclonedgene43ofbacteriophageT4. |
UnitDefinition | Oneunitisdefinedastheamountofenzymethatwillincorporate10nmolofdNTPintoacid-precipitablematerialin30minutesat37°C. |
Components | - T4DNAPolymerase:3,000U/mlin100mMKPO4,1.0mMDTT,0.1mMEDTA,50%glycerol,pH6.5@25°C
- 10XReactionBuffer:500mMNaCl,100mMTris-HCl,100mMMgCl2,10mMDTT,pH7.9@25°C
|
QualityControl | Theabsenceofendodeoxyribonucleasesconfirmedbyappropriatequalitytests. |
StorageCondition | -20°C |
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