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Lucigen/T4 Polynucleotide Kinase/30061-1/1,000 U

  • 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:

  1. Sambrook,J.andRussell,W.(2001)MolecularCloning:ALaboratoryManual,(3rded.),9.55-9.56,9.68-9.69.
  2. Sambrook,J.andRussell,W.(2001)MolecularCloning:ALaboratoryManual,(3rded.),13.16-13.17.
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