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Template-independent DNA polymerases.

S Tabor1

  • 1Harvard Medical School, Boston, Massachusetts, USA.

Current Protocols in Molecular Biology
|February 12, 2008
PubMed
Summary
This summary is machine-generated.

Terminal deoxynucleotidyl transferase (TdT) incorporates deoxynucleotides into DNA 3' termini. This enzyme facilitates DNA fragment cloning, labeling, and synthesis applications.

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Area of Science:

  • Molecular Biology
  • Enzymology

Background:

  • Terminal deoxynucleotidyl transferase (TdT) is a key enzyme in DNA metabolism.
  • Its activity involves the addition of deoxynucleotides to DNA termini.

Purpose of the Study:

  • To describe the reaction conditions for TdT.
  • To outline various applications of TdT in molecular biology.

Main Methods:

  • Purification of TdT from calf thymus.
  • Characterization of enzyme activity under specific reaction conditions.

Main Results:

  • TdT catalyzes the incorporation of deoxynucleotides to the 3'-hydroxyl termini of DNA.
  • Inorganic phosphate is released as a byproduct.
  • Successful application in DNA fragment cloning and labeling.

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Conclusions:

  • TdT is a versatile enzyme with significant applications in DNA manipulation.
  • Detailed reaction conditions enable efficient use of TdT for various molecular biology techniques.