- RecombinantT4PolynucleotideKinase(T4PNK).
- Applicationsincludeend-labelingandinsertpreparationforcloning.
T4PolynucleotideKinasecatalyzesthetransferandexchangeofgamma-phosphateofATPtothe5´-hydroxylterminusofpolynucleotides(double-andsingle-strandedDNAandRNA)andnucleoside3´-monophosphates.
Source:
PurifiedfromastrainofE.colithatcarriesaT4PolynucleotideKinaseexpressionplasmid.
Applications:
- End-labelingDNAforprobesandDNAsequencing(1)
- Additionof5´-phosphatestooligonucleotidestoallowsubsequentligation(2)
Heatinactivation:70°Cfor15minutes.
Storage:
Enzymeandbuffershouldbestoredat-20°C.Enzymeisstablefor12monthsifhandledproperly.
AdditionalInformation:
- AmmoniumionsarepotentinhibitorsofT4PolynucleotideKinase. DNAshouldnotbeprecipitatedfromordissolvedinbufferscontainingammoniumsalts.
- Protruding5’-overhangsaremoreefficientlyphosphorylatedthanblunt-orrecessed5’overhangs.
References:
- Sambrook,J.andRussell,W.(2001)MolecularCloning:ALaboratoryManual,(3rded.),9.55-9.56,9.68-9.69.
- Sambrook,J.andRussell,W.(2001)MolecularCloning:ALaboratoryManual,(3rded.),13.16-13.17.