Molecular Techniques and Methods

Terminal Deoxynucleotidyl Transferase (TdT)

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Terminal Deoxynucleotidyl Transferase, immunoaffinity purified from calf thymus, catalyzes the repetitive addition of mononucleotides from a deoxynucleotide triphosphate to the terminal 3'-OH of a DNA initiator. Single-stranded DNA is preferred as an initiator.




5 X Reaction Buffer
500 mM Cacodylate buffer (pH 6.8)
5 mM CoCl2
0.5 mM DTT
  • The addition of 1 mM Co2+ (as CoCl2) in the Reaction Buffer allows the tailing of any type of 3'-end.




    REFERENCES
  • Kato, K., et al. (1967) J. Biol. Chem. 242: 2780.

  • Okayama, H, Berg, P (1982) Mol. Cell. Biol. 2: 161.

  • Heideker, G, Messing, J (1983) Nucl. Acids Res. 11: 4891.

  • Sambrook, J, Fritsch, FF, Maniatis, T (1989) Molecular Cloning. A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.



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